胸腺瘤组织Slug、TdT、SOX9蛋白表达与临床病理特征及预后的关系分析

摘要 目的：分析锌指转录因子（Slug）、末端脱氧核糖核苷酸转移酶（TdT）、Y染色体性别决定区相关高速泳动族框因子9（SOX9）蛋白表达与胸腺瘤患者临床病理特征及预后的关系。方法：选择2016年5月至2018年11月在本院接受手术切除的113例胸腺瘤组织及癌旁组织,利用免疫组织化学法测定Slug、TdT、SOX9蛋白阳性表达,分析Slug、TdT、SOX9蛋白表达与胸腺瘤患者临床病理特征、预后的关系。结果：胸腺瘤组织Slug、TdT、SOX9蛋白阳性表达率均高于癌旁组织（P＜0.05）;胸腺瘤组织Slug、TdT、SOX9蛋白阳性表达在患者年龄、性别、肿瘤直径、是否合并重症肌无力、是否伴有大血管侵犯方面比较,差异无统计学意义（P＞0.05）;胸腺瘤组织Slug、TdT、SOX9蛋白阳性表达在Masaoka-Koga分期、WHO分型方面比较,差异有统计学意义（P＜0.05）;胸腺瘤组织Slug、TdT、SOX9蛋白阳性表达与Masaoka-Koga分期、WHO分型呈正相关（P＜0.05）;113例胸腺瘤患者随访5年总生存率为81.42%（92/113）,Slug阳性组5年生存率低于Slug阴性组（70.18% VS 92.86%）,TdT阳性组5年生存率低于TdT阴性组（79.790% VS 89.57%）,SOX9阳性组5年生存率低于SOX9阴性组（79.35% VS 90.48%）（χ²=9.669、9.515、17.008,P＜0.05）;Cox比例风险回归模型分析发现,Masaoka-Koga分期（Ⅲ期/Ⅳ期）、WHO分型（B2/B3）、Slug蛋白阳性表达、TdT蛋白阳性表达、SOX9蛋白阳性表达是影响胸腺瘤患者预后的独立危险因素（HR:3.518、2.921、4.536、1.402、2.921,P＜0.05）。结论：胸腺瘤组织Slug、TdT、SOX9呈高表达,三者蛋白表达与患者Masaoka-Koga分期、WHO分型有关,且是影响胸腺瘤患者5年生存率的独立危险因素。     

MYH9: structure,functions and role of non-muscle myosin ⅡA in human disease

MYH9-related diseases (MYH9-RD) are a group of autosomal dominant diseases caused by mutations in the MYH9 gene, which are featured by thrombocytopenia, giant platelets and granulocyte cytoplasmic inclusion bodies. MYH9-RD patients generally suffer from bleeding syndromes, progressive kidney disease, deafness, or cataracts. Here, we reported on a case of MYH9-RD. A novel heterozygous mutation of MYH9 (c.2344-2345delGTinsTA, p.T782Y) was discovered by targeted sequencing technology. Immunofluorescence analysis of neutrophils confirmed abnormal aggregation of MYH9 protein. The results of this study should expand the MYH9 gene mutation spectrum and provide reference for subsequent researchers and genetic counseling.     

CD38基因敲除的淋巴瘤细胞株构建

摘要 目的：构建Luc⁺CD38^-的Raji细胞株,并进行功能的初步验证,为后期探索淋巴瘤细胞CD38位点免疫逃逸现象奠定基础。方法：通过CRISPR-cas9技术和PiggyBac(PB)转座子系统,对Luc⁺Raji细胞的CD38基因位点进行敲除,构建Luc⁺CD38^-Raji细胞株,使用流式细胞术检测与Luc⁺CD38^-Raji细胞株以1：1的比例共孵育CD19 CAR-T和CD38 CAR-T以及未转导的原始T细胞表面活化因子CD69的表达水平,荧光素酶检测法检测上述几组效应细胞对Luc⁺CD38^-Raji细胞株的杀伤效率。结果：成功构建Luc⁺CD38^-Raji细胞,激活实验结果显示,CD19 CAR-T与CD38 CAR-T均可以被Luc⁺Raji细胞激活。而Luc⁺CD38^-Raji19号单克隆细胞由于缺失CD38的表达,仅能够激活CD19 CAR-T。杀伤实验结果显示,两种CAR-T细胞均能够对Luc⁺Raji细胞进行杀伤,而CD38 CAR-T对Luc⁺CD38^-Raji19号单克隆细胞的杀伤效率与原始的T细胞相似。结论：成功构建了Luc⁺CD38^-Raji细胞株,为后期探索淋巴瘤CD38位点免疫逃逸现象奠定基础。     

Physcomitrella patens has lipoxygenases for both eicosanoid and octadecanoid pathways

Mosses have substantial amounts of long chain C20 polyunsaturated fatty acids, such as arachidonic and eicosapentaenoic acid, in addition to the shorter chain C18 α-linolenic and linoleic acids, which are typical substrates of lipoxygenases in flowering plants. To identify the fatty acid substrates used by moss lipoxygenases, eight lipoxygenase genes from Physcomitrella patens were heterologously expressed in Escherichia coli, and then analyzed for lipoxygenase activity using linoleic, α-linolenic and arachidonic acids as substrates. Among the eight moss lipoxygenases, only seven were found to be enzymatically active in vitro, two of which selectively used arachidonic acid as the substrate, while the other five preferred α-linolenic acid. Based on enzyme assays using a Clark-type oxygen electrode, all of the active lipoxygenases had an optimum pH at 7.0, except for one with highest activity at pH 5.0. HPLC analyses indicated that the two arachidonic acid lipoxygenases form (12S)-hydroperoxy eicosatetraenoic acid as the main product, while the other five lipoxygenases produce mainly (13S)-hydroperoxy octadecatrienoic acid from α-linolenic acid. These results suggest that mosses may have both C20 and C18 based oxylipin pathways.     

Molecular Modeling of Calixarenes with Group I Metal Ions

Molecular mechanics calculations have been used to model the geometries of the complexes of Group I metal ions with calix[n]arenes (n = 4,5). A simple procedure in which the calixarene atoms are assigned partial charges on the basis of AM1 calculations and the metal ions are allowed to bind electrostatically to the calixarenes produces surprising good results when the resulting structures are compared to known crystallographic data on the complexes. Encapsulated solvent molecules and/or counterions can be included in the calculations and, indeed, are necessary to reproduce the X-ray data. Electronic Supplementary Material available.     

Compromised proteasome degradation elevates neuronal nitric oxide synthase levels and induces apoptotic cell death

The significance of impairment of proteasome activity in PC12 cells was examined in connection with nitrative/nitrosative stress and apoptotic cell death. Treatment of differentiated PC12 cells with MG132, a proteasome inhibitor, elicited a dose- and time-dependent increase in neuronal nitric oxide synthase (nNOS) protein levels, decreased cell viability, and increased cytotoxicity. Viability and cytotoxicity were ameliorated by L-NAME (a broad NOS inhibitor). Nitric oxide/peroxynitrite formation was increased upon treatment of PC12 cells with MG132 and decreased upon treatment with the combination of MG132 and 7-NI (a specific inhibitor of nNOS). The decreases in cell viability appeared to be effected by an activation of JNK and its effect on mitochondrial Bcl-x_L phosphorylation. These effects are strengthened by the activation of caspase-9 along with increased caspase-3 activity upon treatment of PC12 cells with MG132. These results suggest that impairment of proteasome activity and consequent increases in nNOS levels lead to a nitrative stress that involves the coordinated response of JNK cytosolic signaling and mitochondrion-driven apoptotic pathways.     

Structural basis of the honey bee PBP pheromone and pH-induced conformational change

The behavior of insects and their perception of their surroundings are driven, in a large part, by odorants and pheromones. This is especially true for social insects, such as the honey bee, where the queen controls the development and the caste status of the other individuals. Pheromone perception is a complex phenomenon relying on a cascade of recognition events, initiated in antennae by pheromone recognition by a pheromone-binding protein and finishing with signal transduction at the axon membrane level. With to the objective of deciphering this initial step, we have determined the structures of the bee antennal pheromone-binding protein (ASP1) in the apo form and in complex with the main component of the queen mandibular pheromonal mixture, 9-keto-2(E)-decenoic acid (9-ODA) and with nonpheromonal components. In the apo protein, the C terminus obstructs the binding site. In contrast, ASP1 complexes have different open conformations, depending on the ligand shape, leading to different volumes of the binding cavity. The binding site integrity depends on the C terminus (111-119) conformation, which involves the interplay of two factors; i.e. the presence of a ligand and a low pH. Ligand binding to ASP1 is favored by low pH, opposite to what is observed with other pheromone-binding proteins, such as those of Bombyx mori and Anopheles gambiae.     

Synthesis of cyclooctapeptides: constraints analogues of the peptidic neurotoxin, ω‐agatoxine IVB—an experimental point of view

ω‐AGA IVB is an important lead structure when considering the design of effectors of glutamate release inducting P/Q‐type calcium channels. The best route to achieve the analogues possessing the three‐dimensional arrangement corresponding to the native binding loop was the introduction of constraint by ring formation via side chain to side chain lactamization for suitably protected Lys and Glu residues. Since tryptophane residue located at position 14 of this neuropeptide has been suggested as essential for binding, analogues in which this amino acid was replaced by aza‐tryptophane and alanine were synthesized. The synthesis was carried out on various acid‐labile resins (BARLOS chlorotrityl, Rink amide, PEG‐based or Wang resins), by Fmoc strategy. In this paper, we describe optimization of the peptide cyclization with various protecting groups, and on resin or in solution cyclization experimental parameters. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd.     

Viral tegument proteins restrict cGAS-DNA phase separation to mediate immune evasion

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