壳聚糖酶结构与功能的研究进展

壳聚糖酶是一类对壳聚糖具有较高催化活性而几乎不水解几丁质的糖苷水解酶,其可将高分子量的壳聚糖转化为低分子量的功能性壳寡糖。近年来,对壳聚糖酶的相关研究取得了显著进展,因此,本文对其生化性质、晶体结构、催化机制和蛋白质工程改造进行总结和探讨,并对酶法制备壳寡糖纯品进行展望,这将加深研究者对壳聚糖酶作用机制的认识,推动壳聚糖酶的工业应用。     

嵌合抗原受体T细胞治疗的现状与展望

肿瘤严重威胁着人类健康,当前肿瘤传统的治疗方法有手术治疗、化疗、放疗和靶向药物治疗等。近年来,肿瘤免疫治疗,尤其是嵌合抗原受体（chimeric antigen receptor,CAR） T细胞免疫疗法在基础研究与临床应用中蓬勃发展,并在治疗血液系统恶性肿瘤方面取得了巨大成功。然而,大量研究显示,细胞免疫治疗后可出现不同程度的毒副反应,且部分患者缓解后再次复发。因此,了解细胞治疗面临的挑战与局限性,寻找解决的办法,对继续发挥细胞免疫疗法的潜能具有重要意义。本文就免疫细胞的CAR结构、病毒载体的选择、细胞治疗面临的挑战及前景进行综述。     

基于转录组测序的细风轮花青素合成途径及关键酶基因分析

为进一步理解细风轮花青素合成途径,本研究利用华大基因BGISEQ-500平台对细风轮中的根、茎、叶、花4个组织进行了转录组测序,从头组装后得到128 856个Unigene。KEGG通路表明有40个Unigene编码了细风轮的花青素生物合成途径中6个关键酶。我们对其中的关键酶DFR（二氢黄酮醇还原酶）进行同源比对和空间结构模拟,结果显示DFR序列和结构均具有良好的保守性,且具有高度保守的NAD⁺结合位点,其二级结构主要由α螺旋和β折叠组成,在空间上α螺旋包裹着β折叠,形成“夹心饼干”样结构。     

玉米茎秆强度形成与木质素积累关系的研究

为探究玉米基部节间质量性状与茎秆强度形成的内在关系,该研究选用不同耐密性玉米品种为材料,采用随机区组设计,在田间条件下研究玉米基部节间形态特征、干物质积累的变化特点,分析茎秆内部木质素积累动态变化及其相关合成酶活性对茎秆强度形成的影响。结果表明：(1)耐密品种‘先玉335’基部节间单位长度干重(DWUL)和直径均较高,不同品种的茎秆强度快速形成时期有一定差异,与木质素的积累密切相关。(2)耐密品种茎秆穿刺强度(RPS)和木质素积累快速形成时期较不耐密品种‘新玉41’长5~7 d,穿刺强度高于不耐密品种24.9%~36.6%,其木质素积累量高于不耐密品种12.5%~47.0%,且RPS和木质素积累速率较不耐密品种快。(3)玉米抽雄期(VT)前是基部节间木质素快速积累的关键时期,玉米大喇叭口期(V12~V15)酶活性与抽雄期木质素积累量呈显著或极显著正相关,对茎秆强度形成至关重要。(4)在玉米12叶期耐密品种‘先玉335’的木质素合成相关酶均显著高于不耐密品种‘新玉41’,PAL、TAL、CAD和POD分别较‘新玉41’高1.85、0.30、0.11和0.42 U·mg^-1。研究认为,玉米大喇叭口期茎秆干物质积累量较高、木质素合成相关酶的活性较强,能有效促进木质素的快速积累,增加茎秆抗倒伏强度,进而提高玉米茎秆抗倒伏能力。     

混合接种球孢白僵菌与摩西球囊霉对烟草促生抗逆影响

利用球孢白僵菌Beauveria bassiana与摩西球囊霉菌Glomus mosseae以单独和混合接种的方式在烟草Nicotiana tabacum苗期进行接种处理,检测烟草苗期和成熟期的农艺性状和生理特性。结果表明：在育苗期,混合接种处理烟苗具有最大的叶长和最大叶宽,与对照相比提高了30.27%和42.98%,球孢白僵菌处理组生物量最大,AM真菌处理组烟草株高最高;在成熟期,混合接种处理组烟草的株高、最大叶长、最大叶宽、茎围、生物量等指标均优于单接种处理,且过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、过氧化物酶（POD）等抗氧化酶活性最高,丙二醛（MDA）含量最低。球孢白僵菌和AM真菌可协同提高烟草生长和抗逆能力。     

Construction of a robust prognostic model for adult adrenocortical carcinoma: Results from bioinformatics and real-world data

This study aims to construct a robust prognostic model for adult adrenocortical carcinoma (ACC) by large-scale multiomics analysis and real-world data. The RPPA data, gene expression profiles and clinical information of adult ACC patients were obtained from The Cancer Proteome Atlas (TCPA), Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Integrated prognosis-related proteins (IPRPs) model was constructed. Immunohistochemistry was used to validate the prognostic value of the IPRPs model in Fudan University Shanghai Cancer Center (FUSCC) cohort. 76 ACC cases from TCGA and 22 ACC cases from GSE10927 in NCBI’s GEO database with full data for clinical information and gene expression were utilized to validate the effectiveness of the IPRPs model. Higher FASN (P = .039), FIBRONECTIN (P < .001), TFRC (P < .001), TSC1 (P < .001) expression indicated significantly worse overall survival for adult ACC patients. Risk assessment suggested significantly a strong predictive capacity of IPRPs model for poor overall survival (P < .05). IPRPs model showed a little stronger ability for predicting prognosis than Ki-67 protein in FUSCC cohort (P = .003, HR = 3.947; P = .005, HR = 3.787). In external validation of IPRPs model using gene expression data, IPRPs model showed strong ability for predicting prognosis in TCGA cohort (P = .005, HR = 3.061) and it exhibited best ability for predicting prognosis in GSE10927 cohort (P = .0898, HR = 2.318). This research constructed IPRPs model for predicting adult ACC patients’ prognosis using proteomic data, gene expression data and real-world data and this prognostic model showed stronger predictive value than other biomarkers (Ki-67, Beta-catenin, etc) in multi-cohorts.     

Nox和氧化应激与糖尿病

Nox(phagocyte-like NADPH oxidase)是吞噬细胞NADPH氧化酶催化亚基 gp91phox的一系列同源物,广泛分布于体内多种非吞噬细胞.与NADPH氧化酶类似, Nox激活后可产生ROS,Nox产生的ROS是线粒体外ROS的主要来源.Nox产生的ROS,在控制新陈代谢,调节葡萄糖刺激的胰岛素分泌(glucose-stimulated insulin secretion,GSIS),促使胰岛β细胞凋亡、胰岛功能障碍和糖尿病及其并发症的发 生、发展中,发挥着重要作用.调节Nox的活性,改善机体内氧化应激水平,有望成为治疗糖尿病及其并发症的有效新途径.     

以整合素αvβ3为靶点的RGD配体在抗肿瘤血管新生中的应用

整合素αvβ3是一种能特异性识别RGD序列的膜受体蛋白,其与含RGD（Arg-Gly-Asp）模体的蛋白质结合的特异性在肿瘤细胞的粘附、迁移、浸润及肿瘤血管新生中起重要作用.由于整合素αvβ3在多种肿瘤细胞表面高表达而在正常细胞中低表达或不表达,因此其成为肿瘤治疗的理想靶点.肿瘤新生血管为肿瘤的生长提供营养,因此近年来抑制肿瘤血管新生也成为肿瘤治疗的重要途径.有研究显示,几种RGD毒素蛋白不但以整合素αvβ3为靶点靶向结合到肿瘤部位从而具有直接抗肿瘤细胞增殖、黏附、迁移及浸润功能,而且它们还具有抗肿瘤血管新生的作用,因此RGD毒素蛋白可从上述两方面抑制肿瘤生长与转移.本文就整合素αvβ3为靶向的RGD配体结构特点及其在肿瘤治疗中的靶向治疗和抗血管新生应用及前景加以综述.     

Metabolic engineering of Corynebacterium glutamicum for increasing the production of l-ornithine by increasing NADPH availability

The experiments presented here were based on the conclusions of our previous proteomic analysis. Increasing the availability of glutamate by overexpression of the genes encoding enzymes in the l-ornithine biosynthesis pathway upstream of glutamate and disruption of speE, which encodes spermidine synthase, improved l-ornithine production by Corynebacterium glutamicum. Production of l-ornithine requires 2 moles of NADPH per mole of l-ornithine. Thus, the effect of NADPH availability on l-ornithine production was also investigated. Expression of Clostridium acetobutylicum gapC, which encodes NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, and Bacillus subtilis rocG, which encodes NAD-dependent glutamate dehydrogenase, led to an increase of l-ornithine concentration caused by greater availability of NADPH. Quantitative real-time PCR analysis demonstrates that the increased levels of NADPH resulted from the expression of the gapC or rocG gene rather than that of genes (gnd, icd, and ppnK) involved in NADPH biosynthesis. The resulting strain, C. glutamicum ΔAPRE::rocG, produced 14.84 g l^?1 of l-ornithine. This strategy of overexpression of gapC and rocG will be useful for improving production of target compounds using NADPH as reducing equivalent within their synthetic pathways.     

Preventive Effect of Halofuginone on Concanavalin A-Induced Liver Fibrosis

Halofuginone (HF) is an active component of extracts derived from the plant alkaloid febrifugine and has shown therapeutic promise in animal models of fibrotic disease. Our main objectives were to clarify the suppressive effect of HF on concanavalin A (ConA)-induced liver fibrosis. ConA injection into the tail vein caused a great increase in the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, while orally administration of HF significantly decreased the levels of the transaminases. In addition, the levels of hyaluronic acid (HA), procollagen III (PCIII) and TGF-β1 in the serum and collagen I, α-SMA, tissue inhibitors of metalloproteinase 2 (TIMP2) and Smad3 in the liver tissue were significantly lowered with the treatment of HF. Histological examination also demonstrated that HF significantly reduced the severity of liver fibrosis. Since ConA-induced liver fibrosis is caused by the repeated activation of T cells, immunomodulatory substances might be responsible for the suppressive effect of HF. We found that the production of nuclear factor (NF)-kB in the serum was increased in ConA-treated group, while decreased significantly with the treatment of HF. The changes of inflammatory cytokines tumor necrosis factor (TNF-α), IL-6 and IL-1β in the serum followed the same rhythm. All together, our findings indicate that orally administration HF (10ppm) would attenuate the liver fibrosis by suppressing the synthesis of collagen I and inflammation-mediated liver injury.