中欧组织间合作研究项目MIX-UP助力实现“碳中和”

随着全球塑料循环体系的变革升级,提高塑料的回收利用不仅可以减少塑料在生命周期中的碳排放,还可以解决废塑料潜在的生态环境危害。文中介绍了2019年国家自然科学基金组织间国际 (地区) 合作研究项目“废塑料资源高效生物降解转化的关键科学问题与技术 (MIXed plastics biodegradation and UPcycling using microbial communities,MIX-UP)”。该项目聚焦“塑料污染”这一全球化的问题,围绕中欧双方确定的“塑料生物降解菌群”研究领域,联合中欧双方14家优势科研单位,开展实质性的重大前沿合作研究。针对废塑料生物降解中存在的解聚与重塑两个难题,项目以难降解石油基塑料 (PP、PE、PUR、PET和PS) 以及生物可降解塑料 (PLA和PHA) 的混合废塑料作为研究对象,从塑料微生物降解途径解析及关键元件的挖掘与改造、塑料高效降解混菌/多酶体系的构建与功能调控、塑料降解物的高值化炼制途径设计与利用策略3个方面展开研究。本项目将突破废塑料生物降解转化中高效降解元件挖掘、塑料降解物高值化利用的关键科学问题与技术,探索一条废塑料资源化、高值化、循环化、低碳化的新塑料循环路线,建立以“降塑再造”为核心理念的废塑料生物炼制体系,丰富我国固废资源化生物技术利用平台。项目的实施不仅有助于提升我国塑料 (生物) 循环经济的理论基础和关键技术水平,还可以推动我国与国际科研院所的多边交流与合作,促进我国在生物技术领域的创新发展,助力我国碳中和目标的实现。     

海南地区与环渤海湾美人鱼发光杆菌美人鱼亚种水产动物分离株的表型与遗传特征分析

美人鱼发光杆菌美人鱼亚种（Photobacterium damselae subsp.damselae,PDD）是一种广泛分布于海洋环境内的重要病原菌,可导致多种海洋生物患病死亡,近年来在我国不同养殖区域内均有发现和报道。[目的] 通过生理代谢表型、毒力基因分布和分子遗传分析,系统比较我国海南地区和环渤海湾不同株系PDD的表型与遗传多样性特征。[方法] 使用选择性培养基测定PDD的蔗糖发酵能力、运动性、溶血性和磷脂酶活性;通过PCR检测质粒pPHDD1复制起始点基因pPHDD1 replication origin和毒力基因dly,hlyA_pl,hlyA_ch,plpV;基于toxR基因序列构建系统发育树,分析16株PDD的遗传关系。[结果] 4株PDD蔗糖发酵测试呈阳性,12株PDD蔗糖发酵测试呈阴性。不同株系PDD运动能力存在显著差异,其中环渤海湾分离株Pdd1608、Pdd2009和Pdd1704的运动能力较强。仅有环渤海湾分离株Pdd1608表现为强溶血性、强磷脂酶活性;海南地区分离株Pdd1612和Pdd0912表现为弱溶血性、无磷脂酶活性;其余菌株表现为弱溶血性、弱磷脂酶活性。菌株Pdd1608的毒力基因谱为pPHDD1 replication origin-dly-hlyA_p-hlyA_ch-plpV,其余菌株的毒力基因谱为hlyA_ch-plpV。系统发育分析揭示不同株系PDD具有高度的遗传多样性。[结论] 流行于海南地区和环渤海湾的PDD可能都由多克隆种群组成,它们具有复杂的表型特征、相似的毒力基因分布和高度的遗传多样性。     

屯昌猪MYLPF基因序列分析及其组织表达量检测

为获得屯昌猪MYLPF基因的CDS区序列并分析其分子结构特征,研究屯昌猪、杜洛克猪以及其杂交F1代猪不同组织中的MYLPF mRNA表达水平,本研究以GenBank上公布的猪MYLPF基因序列(登录号:NM_001006592)为参考设计引物,通过RT-PCR扩增、测序获得屯昌猪MYLPF基因CDS区.结果显示:该基因CDS区长度510 bp,编码169个氨基酸,在CDS区存在33 G＞T的同义突变;该基因编码产物既没有跨膜结构,也不存在信号肽,属于非分泌型蛋白,主要在细胞质中发挥作用,预测存在1个潜在的糖基化位点和9个磷酸化位点,α螺旋区域在预测的二级结构中占比最大.荧光定量PCR检测结果显示MYLPF基因在屯昌猪背最长肌中表达量最高,屯昌猪背最长肌中MYLPF mRNA的表达量均显著高于杜洛克猪及其杂交F1代猪.本研究为探明MYLPF基因对猪肌内脂肪的影响提供了分子理论依据.     

Blocking of the PD-1/PD-L1 interaction by a novel cyclic peptide inhibitor for cancer immunotherapy

The interaction of PD-1/PD-L1 allows tumor cells to escape from immune surveillance. Clinical success of the antibody drugs has proven that blockade of PD-1/PD-L1 pathway is a promising strategy for cancer immunotherapy. Here, we developed a cyclic peptide C8 by using Ph.D.-C7 C phage display technology. C8 showed high binding affinity with h PD-1 and could effectively interfere the interaction of PD-1/PD-L1. Furthermore, C8 could stimulate CD8+T cell activation in human peripheral blood mononuclear cells(PBMCs). We also observed that C8 could suppress tumor growth in CT26 and B16-OVA, as well as anti-PD-1 antibody resistant B16 mouse model. CD8+T cells infiltration significantly increased in tumor microenvironment, and IFN-γ secretion by CD8+T cells in draining lymph nodes also increased. Simultaneously, we exploited T cells depletion models and confirmed that C8 exerted anti-tumor effects via activating CD8+T cells dependent manner. The interaction model of C8 with h PD-1 was simulated and confirmed by alanine scanning. In conclusion, C8 shows anti-tumor capability by blockade of PD-1/PD-L1 interaction, and C8 may provide an alternative candidate for cancer immunotherapy.     

黄山典型植被类型土壤真菌群落特征及其影响因素

黄山地势高差明显,植被类型多样,生态系统保存完整,是研究森林生态系统土壤真菌群落的天然实验室。本研究采集黄山典型植被下土壤样本,利用Illumina NovaSeq高通量测序技术分析土壤真菌群落结构,结合土壤理化性质探讨不同植被类型影响真菌群落组成的潜在因素。结果共检测到13个真菌门,优势真菌门依次为：担子菌门Basidiomycota,获得38目,202属,相对丰度介于7.30%-90.71%,在常绿落叶阔叶混交林、山地矮林及落叶阔叶林中出现高值,局部呈现先增后减的单峰变化格局;子囊菌门Ascomycota有56目,393属,相对丰度介于4.69%-53.07%,随着典型植被类型变化无明显变化规律;被孢霉门Mortierellomycota获得1目和2属,相对丰度介于2.88%-29.92%,随着典型植被类型变化呈现U型变化模式;5种植被类型土壤中共检测到34个不同分类单元的真菌指示类群,落叶阔叶林土壤真菌指示类群最为丰富,占67%;pH显著影响土壤真菌α多样性（Pearson,P<0.001）,是黄山土壤真菌群落变异的主控因子（Monte Corlo 检验,P<0.01）。     

不同胃癌前病变中医证型PG、G-17、CEA和叶酸水平变化及相关性研究

目的:对不同胃癌前病变中医证型和血清胃蛋白酶原(pepsinogen,PG)、胃泌素-17(gastrin-17,G-17)、癌胚抗原(carcinoembryonic antigen,CEA)和叶酸水平变化的关系进行探讨,为胃癌前病变的诊断提供一定的依据。方法:以80例胃癌前病变(precancerous lesion of gastric cancer,PLGC)患者研究组,80例健康者为对照组,对研究组患者进行中医临床辨证分型,对两组研究对象的血清PG、G-17、CEA和叶酸进行测定比较。结果:研究组PLGC患者中医证型分布不均匀,差异显著(P<0.05),由多到少依次为湿热蕴胃并/兼脾胃虚寒证>胃络瘀阻并/兼气阴两虚证>痰湿中阻并/兼脾胃气虚证>肝胃气滞并/兼气阴两虚证>肝胃气滞并/兼脾胃虚寒证>湿热蕴胃并/兼胃阴不足证。PLGC不同中医证型患者血清PG I和PG II水平差异显著(P<0.05);与对照组比较,各证型PG I水平均显著降低,PG II水平均显著升高(P<0.05)。且湿热蕴胃并/兼脾胃虚寒证和胃络瘀阻并/兼气阴两虚证的PG I水平显著低于其他证候,血清PG II水平显著高于其他(P<0.05)。与对照组比较,研究组不同证候的G-17、CEA水平显著升高,叶酸水平显著降低(P<0.05);观察组中湿热蕴胃并/兼脾胃虚寒证和胃络瘀阻并/兼气阴两虚证G-17、CEA显著高于其他证候,叶酸水平显著低于其他证候(P<0.05)。结论:胃癌前病变不同中医证型血清PG、G-17、CEA和叶酸存在明显差异。     

长链非编码RNA KCNQ1OT1影响脂多糖诱导的血管内皮细胞凋亡及其炎性因子表达的机制

该研究探讨了长链非编码RNA KCNQ1OT1对脂多糖(LPS)诱导的血管内皮细胞(VEC)凋亡和炎性因子表达的影响以及其可能机制。通过体外培养VEC,分别转染KCNQ1OT1过表达载体、miR-223抑制剂或共转染KCNQ1OT1过表达载体与miR-223模拟物后,用1.0 mg/mL LPS干预24 h,然后采用RT-qPCR法检测细胞中KCNQ1OT1和miR-223的表达水平,流式细胞仪检测细胞凋亡,Western blot检测细胞中Bcl-2和Bax蛋白表达,ELISA试剂盒检测细胞培养上清中TNF-α、IL-1和IL-6水平。双荧光素酶报告基因实验验证KCNQ1OT1与miR-223的调控关系。结果显示,LPS可抑制VEC中KCNQ1OT1的表达,而促进miR-223表达;上调KCNQ1OT1或下调miR-223后均可降低LPS诱导的VEC凋亡率、Bax蛋白及TNF-α、IL-1和IL-6表达(P<0.05),而促进Bcl-2蛋白表达(P<0.05)。KCNQ1OT1靶向负调控miR-223表达,上调miR-223则逆转上调KCNQ1OT1对LPS诱导的VEC凋亡及炎性因子表达的抑制作用。这表明,上调KCNQ1OT1抑制LPS诱导的VEC凋亡及炎性因子表达,其作用机制可能与靶向负调控miR-223有关,KCNQ1OT1/miR-223轴可能为血管内皮细胞损伤的治疗提供了新靶点。     

Photobiomodulation suppresses JNK3 by activation of ERK/MKP7 to attenuate AMPA receptor endocytosis in Alzheimer's disease

Alzheimer's disease (AD), a severe age‐related neurodegenerative disorder, lacks effective therapeutic methods at present. Physical approaches such as gamma frequency light flicker that can effectively reduce amyloid load have been reported recently. Our previous research showed that a physical method named photobiomodulation (PBM) therapy rescues Aβ‐induced dendritic atrophy in vitro. However, it remains to be further investigated the mechanism by which PBM affects AD‐related multiple pathological features to improve learning and memory deficits. Here, we found that PBM attenuated Aβ‐induced synaptic dysfunction and neuronal death through MKP7‐dependent suppression of JNK3, a brain‐specific JNK isoform related to neurodegeneration. The results showed PBM‐attenuated amyloid load, AMPA receptor endocytosis, dendrite injury, and inflammatory responses, thereby rescuing memory deficits in APP/PS1 mice. We noted JNK3 phosphorylation was dramatically decreased after PBM treatment in vivo and in vitro. Mechanistically, PBM activated ERK, which subsequently phosphorylated and stabilized MKP7, resulting in JNK3 inactivation. Furthermore, activation of ERK/MKP7 signaling by PBM increased the level of AMPA receptor subunit GluR 1 phosphorylation and attenuated AMPA receptor endocytosis in an AD pathological model. Collectively, these data demonstrated that PBM has potential therapeutic value in reducing multiple pathological features associated with AD, which is achieved by regulating JNK3, thus providing a noninvasive, and drug‐free therapeutic strategy to impede AD progression.     

The herpesvirus accessory protein γ134.5 facilitates viral replication by disabling mitochondrial translocation of RIG-I

RIG-I and MDA5 are cytoplasmic RNA sensors that mediate cell intrinsic immunity against viral pathogens. While it has been well-established that RIG-I and MDA5 recognize RNA viruses, their interactive network with DNA viruses, including herpes simplex virus 1 (HSV-1), remains less clear. Using a combination of RNA-deep sequencing and genetic studies, we show that the γ₁34.5 gene product, a virus-encoded virulence factor, enables HSV growth by neutralization of RIG-I dependent restriction. When expressed in mammalian cells, HSV-1 γ₁34.5 targets RIG-I, which cripples cytosolic RNA sensing and subsequently suppresses antiviral gene expression. Rather than inhibition of RIG-I K63-linked ubiquitination, the γ₁34.5 protein precludes the assembly of RIG-I and cellular chaperone 14-3-3ε into an active complex for mitochondrial translocation. The γ₁34.5-mediated inhibition of RIG-I-14-3-3ε binding abrogates the access of RIG-I to mitochondrial antiviral-signaling protein (MAVS) and activation of interferon regulatory factor 3. As such, unlike wild type virus HSV-1, a recombinant HSV-1 in which γ₁34.5 is deleted elicits efficient cytokine induction and replicates poorly, while genetic ablation of RIG-I expression, but not of MDA5 expression, rescues viral growth. Collectively, these findings suggest that viral suppression of cytosolic RNA sensing is a key determinant in the evolutionary arms race of a large DNA virus and its host.