抑制CD36与Nogo-B表达抑制三阴性乳腺癌细胞增殖与迁移

分化聚类36（cluster of differentiation 36,CD36）是一种位于细胞表面的膜蛋白受体,可以结合并转运脂肪酸。内质网膜蛋白4B （Nogo-B）在肝脏中调控脂肪酸代谢而影响肝癌的发展。目前并不清楚CD36和Nogo-B的相互作用是否能够影响乳腺癌细胞的增殖和迁移。本研究在三阴性乳腺癌（triple-negative breast cancer,TNBC）细胞中同时干预CD36与Nogo-B的表达来探索它们对细胞增殖与迁移的影响。结果表明在三阴性乳腺癌细胞中,单独抑制CD36或Nogo-B的表达都能够抑制细胞的增殖与迁移;同时抑制CD36与Nogo-B的表达时,这种抑制效果更加明显,且Vimentin、B细胞淋巴瘤-2（B-cell lympoma-2,BCL2）和增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）的表达受到抑制。在小鼠移植瘤模型中,E0771细胞转染CD36或Nogo-B的siRNA后成瘤能力降低;同时敲减CD36和Nogo-B时,肿瘤生长速度显著减慢。机制研究发现,抑制CD36和Nogo-B表达能够抑制脂肪酸结合蛋白4（fatty acid binding protein 4,FABP4）和脂肪酸转运蛋白4（fatty acid transport protein 4,FATP4） mRNA的含量,同时CD36和Nogo-B过表达刺激的细胞增殖被FABP4的siRNA降低,预示着抑制乳腺癌细胞中CD36与Nogo-B的表达可能通过抑制脂肪酸的吸收和转运而抑制细胞的生长和迁移。此外,抑制CD36与Nogo-B的表达可激活P53-P21-Rb信号通路,参与抑制CD36与Nogo-B表达而抑制的细胞增殖与迁移。本研究证明同时抑制CD36和Nogo-B的表达能够协同抑制三阴性乳腺癌细胞的增殖和迁移,为临床抗三阴性乳腺癌药物的开发提供了新的靶点。     

肺炎支原体感染对川崎病的影响及机制探讨

目的分析肺炎支原体感染对川崎病的影响及其机制,以期指导临床诊疗。方法回顾性分析2013年8月至2018年8月我科住院的496例川崎病患儿临床资料,其中合并肺炎支原体感染组193例,未合并肺炎支原体感染组303例。应用倾向评分匹配法1∶1校正2组的年龄、性别、是否为不完全型川崎病、是否使用糖皮质激素、丙种球蛋白使用时间、丙种球蛋白使用方法和阿司匹林初始剂量,比较2组患儿的总发热天数、冠状动脉直径、冠状动脉扩张发生率、冠状动脉瘤发生率和丙种球蛋白无反应发生率。结果川崎病合并肺炎支原体感染组男性患儿多,更易发生颈部淋巴结肿大,年龄、中性粒细胞百分比、血沉和低密度脂蛋白高于川崎病未合并肺炎支原体感染组,血钾、血钙、高密度脂蛋白和白蛋白低于川崎病未合并肺炎支原体感染组,差异有统计学意义(均P<0.05)。应用倾向评分匹配法1∶1配对后发现,川崎病合并肺炎支原体感染组患儿总发热时间更长[8(6~10)d vs 7(6~9)d,Z=-2.089,P<0.05),冠状动脉直径值更大[(2.81±0.81)mm vs(2.63±0.45)mm,t=2.532,P<0.05],冠状动脉瘤发生率更高(5.5% vs 1.1%,χ²=5.516,P<0.05)。结论肺炎支原体感染可能引起川崎病患儿脂质代谢及电解质的紊乱,其炎症反应更强,持续时间更长,对冠状动脉损伤更大。     

Preparation,cellular uptake and angiogenic suppression of shikonin-containing liposomes in?vitro and in?vivo

Shikonin has anticancer activity, but it has not yet been applied into clinical use. In the present study, shikonin was prepared using liposomes. We aimed to examine several aspects of sh-L (shikonin-containing liposomes): preparation, angiogenic suppression and cellular uptake through self-fluorescence. Sh-L were prepared using soybean phospholipid and cholesterol to form the membrane and shikonin was encapsulated into the phospholipid membrane. Three liposomes were prepared with shikonin. They had red fluorescence and were analysed using a flow cytometer. Angiogenic suppression of sh-L was determined using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide], Transwell tests, chick CAM (chorioallantoic membrane) and Matrigel™ plug assay. MTT assay showed the median IC₅₀ (inhibitory concentrations) as follows: shikonin, sh-L₁ and sh-L₂ were 4.99±0.23, 5.81±0.57 and 7.17±0.69 μM, respectively. The inhibition rates of migration were 53.58±7.05, 46.56±4.36 and 41.19±3.59% for 3.15 μM shikonin, sh-L₁ and sh-L₂, respectively. The results of CAM and Matrigel plug assay demonstrated that shikonin and sh-L can decrease neovascularization. Effect of shikonin was more obvious than sh-L at the same concentration. The results showed that sh-L decreased the toxicity, the rate of inhibition of migration and angiogenic suppression. The cellular uptake of the sh-L could be pictured because of the self-fluorescence. The self-fluorescence will be useful for conducting further research. Sh-L might be an excellent preparation for future clinical application to cancer patients.     

Chinese white truffles shape the ectomycorrhizal microbial communities of Corylus avellana

Here, we investigated the influence of Chinese white truffle (Tuber panzhihuanense) symbioses on the microbial communities associated with Corylus avellana during the early development stage of symbiosis. The microbial communities associated with ectomycorrhizae, and associated with roots without T. panzhihuanense colonization, were determined via high-throughput sequencing of bacterial 16S rRNA genes and fungal ITS genes. Microbial community diversity was higher in the communities associated with the ectomycorrhizae than in the control treatment. Further, bacterial and fungal community structures were different in samples containing T. panzhihuanense in association with C. avellana compared to the control samples. In particular, the bacterial genera Rhizobium, Pedomicrobium, and Herbiconiux were more abundant in the ectomycorrhizae, in addition to the fungal genus Monographella. Moreover, there were clear differences in some physicochemical properties among the rhizosphere soils of the two treatments. Statistical analyses indicated that soil properties including exchangeable magnesium and exchangeable calcium prominently influenced microbial community structure. Lastly, inference of bacterial metabolic functions indicated that sugar and protein metabolism functions were significantly more enriched in the communities associated with the ectomycorrhizae from C. avellana mycorrhized with T. panzhihuanense compared to communities from roots of cultivated C. avellana without T. panzhihuanense. Taken together, these results highlight the interactions among ectomycorrhizal fungi, soil properties, and microbial communities that are associated with host plants and further our understanding of the ecology and cultivation of the economically important T. panzhihuanense truffles.

     

斑马鱼转座子时空表达特性

转座子是基因组中可移动和扩展的元件,能够插入新的位点,影响基因组和基因的结构和功能,是基因组进化的内在驱动。为探讨转座子的时空表达特性,首先通过生物信息学方法鉴定出斑马鱼9个疑似活性转座子,包括DNA转座子Tc1家族(Tc-a、Tc-b、Tc-c、Tc-d、Tc-e)、反转录转座子ERV家族(ERV-1、ERV-2)和LINE家族(L1-323、L1-21),然后采用实时荧光定量PCR法检测上述转座子在斑马鱼早期胚胎发育7个阶段及成鱼各主要脏器的表达活性。结果表明:Tc1家族在0.75、2.00、3.00 h各阶段无转录活性,在6.00、15.00、24.00、48.00 h各阶段各转座子均有较高转录活性;反转录转座子转录活性最早出现于3 h,最晚出现于15 h,且随着发育时间的延长,转录活性显著增强。9种转座子在成鱼心脏、大脑、肌肉、肝脏、睾丸和卵巢均有表达,且大脑和心脏中的表达水平显著高于其他组织,睾丸表达水平最低。分析表明转座子的表达具有时间和组织的特异性,可能参与斑马鱼胚胎和组织器官发育调控,尤其是大脑和心脏发育。这些结果为进一步研究转座子是否具有基因表达调控功能提供重要参考。     

Ageing related thyroid deficiency increases brain-targeted transport of liver-derived ApoE4-laden exosomes leading to cognitive impairment

Alzheimer’s disease (AD) is the prevalent cause of dementia in the ageing world population. Apolipoprotein E4 (ApoE4) allele is the key genetic risk factor for AD, although the mechanisms linking ApoE4 with neurocognitive impairments and aberrant metabolism remains to be fully characterised. We discovered a significant increase in the ApoE4 content of serum exosomes in old healthy subjects and AD patients carrying ApoE4 allele as compared with healthy adults. Elevated exosomal ApoE4 demonstrated significant inverse correlation with serum level of thyroid hormones and cognitive function. We analysed effects of ApoE4-containing peripheral exosomes on neural cells and neurological outputs in aged or thyroidectomised young mice. Ageing-associated hypothyroidism as well as acute thyroidectomy augmented transport of liver-derived ApoE4 reach exosomes into the brain, where ApoE4 activated nucleotide-binding oligomerisation domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome by increasing cholesterol level in neural cells. This, in turn, affected cognition, locomotion and mood. Our study reveals pathological potential of exosomes-mediated relocation of ApoE4 from the periphery to the brain, this process can represent potential therapeutic target.Subject terms: Cognitive neuroscience, Alzheimer''s disease, Cellular neuroscience     

凉山州新银合欢根瘤菌的共生有效性及遗传多样性

【目的】研究分离自四川凉山州新银合欢根瘤菌的遗传多样性和共生有效性。【方法】采用16S rRNA RFLP、BOX-PCR、AFLP、多位点持家基因序列的联合分析及无氮水培法对33株供试新银合欢根瘤菌的遗传多样性和共生有效性进行研究。【结果】分析表明,3种方法在属水平的分群结果具有较好的一致性,有1个Mesorhizobium属的菌株、3个Bradyrhizobium属的菌株、3个Rhizobium属的菌株,26个相似度较高的菌株属Sinorhizobium。16S rRNA-recA-atpD-glnII序列联合构建的新银合欢根瘤菌系统发育树表明,SCAU203、SCAU211可能分别是Rhizobium和Bradyrhizobium的新类群,另外3个代表菌株分别位于Sinorhizobium、Mesorhizobium、Bradyrhizobium分支,分别与S.americanum、M.Plurifarium、R.huautlense亲缘关系最近。无氮水培接种试验筛选出2个共生固氮效果好、与不接种对照处理差异达显著水平的菌株SCAU229和SCAU307,有3个菌株不仅不具共生有效性,甚至不利于宿主的生长,其余84%的供试菌为低效或无效菌株。【结论】凉山州新银合欢根瘤菌具有丰富的遗传多样性,分布于4个属:Rhizobium、Bradyrhizobium、Mesorhizobium、Bradyrhizobium,79%为Sinorhizobium属的菌株,优势菌群为Sinorhizobium。该区的新银合欢根瘤菌大多数的共生有效性差。     

PDCD10/CCM3 acts downstream of {gamma}-protocadherins to regulate neuronal survival

γ-Protocadherins (PCDH-γ) regulate neuronal survival in the vertebrate central nervous system. The molecular mechanisms of how PCDH-γ mediates this function are still not understood. In this study, we show that through their common cytoplasmic domain, different PCDH-γ isoforms interact with an intracellular adaptor protein named PDCD10 (programmed cell death 10). PDCD10 is also known as CCM3, a causative genetic defect for cerebral cavernous malformations in humans. Using RNAi-mediated knockdown, we demonstrate that PDCD10 is required for the occurrence of apoptosis upon PCDH-γ depletion in developing chicken spinal neurons. Moreover, overexpression of PDCD10 is sufficient to induce neuronal apoptosis. Taken together, our data reveal a novel function for PDCD10/CCM3, acting as a critical regulator of neuronal survival during development.     

A built-in mechanism to mitigate the spread of insect-resistance and herbicide-tolerance transgenes into weedy rice populations

Background

The major challenge of cultivating genetically modified (GM) rice (Oryza sativa) at the commercial scale is to prevent the spread of transgenes from GM cultivated rice to its coexisting weedy rice (O. sativa f. spontanea). The strategic development of GM rice with a built-in control mechanism can mitigate transgene spread in weedy rice populations.

Methodology/Principal Findings

An RNAi cassette suppressing the expression of the bentazon detoxifying enzyme CYP81A6 was constructed into the T-DNA which contained two tightly linked transgenes expressing the Bt insecticidal protein Cry1Ab and the glyphosate tolerant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), respectively. GM rice plants developed from this T-DNA were resistant to lepidopteran pests and tolerant to glyphosate, but sensitive to bentazon. The application of bentazon of 2000 mg/L at the rate of 40 mL/m², which is approximately the recommended dose for the field application to control common rice weeds, killed all F₂ plants containing the transgenes generated from the Crop-weed hybrids between a GM rice line (CGH-13) and two weedy rice strains (PI-63 and PI-1401).

Conclusions/Significance

Weedy rice plants containing transgenes from GM rice through gene flow can be selectively killed by the spray of bentazon when a non-GM rice variety is cultivated alternately in a few-year interval. The built-in control mechanism in combination of cropping management is likely to mitigate the spread of transgenes into weedy rice populations.