浙江菌类药资源调查及利用研究初报

本文较全面地了浙江菌类药中药资源。其内容包括开展本项目的方法,研究结果。分别对本省自然环境、野生菌类药中药资源的分布进行了重点论述,简要介绍了菌类药调查结果,重点研究种类,并对本省的菌类药现状和展望作了评述;文后列有本项目研究中公开发表的文献,使本方基本能反映本专题的研究内容。     

HDAC8 drives spindle organization during meiotic maturation of porcine oocytes

ObjectivesHistone deacetylase 8 (HDAC8) is one of the class I HDAC family proteins, which participates in the neuronal disorders, parasitic/viral infections, tumorigenesis and many other biological processes. However, its potential function during female germ cell development has not yet been fully understood.Materials and methodsHDAC8‐targeting siRNA was microinjected into GV oocytes to deplete HDAC8. PCI‐34051 was used to inhibit the enzyme activity of HDAC8. Immunostaining, immunoblotting and fluorescence intensity quantification were applied to assess the effects of HDAC8 depletion or inhibition on the oocyte meiotic maturation, spindle/chromosome structure, γ‐tubulin dynamics and acetylation level of α‐tubulin.ResultsWe observed that HDAC8 was localized in the nucleus at GV stage and then translocated to the spindle apparatus from GVBD to M II stages in porcine oocytes. Depletion of HDAC8 led to the oocyte meiotic failure by showing the reduced polar body extrusion rate. In addition, depletion of HDAC8 resulted in aberrant spindle morphologies and misaligned chromosomes due to the defective recruitment of γ‐tubulin to the spindle poles. Notably, these meiotic defects were photocopied by inhibition of HDAC8 activity using its specific inhibitor PCI‐34051. However, inhibition of HDAC8 did not affect microtubule stability as assessed by the acetylation level of α‐tubulin.ConclusionsCollectively, our findings demonstrate that HDAC8 acts as a regulator of spindle assembly during porcine oocyte meiotic maturation.     

四种细菌L型的形态定量分析

用抗生素或臭氧的方法诱导出四种细菌和Ｌ型后,用图像分析技术定量检测Ｌ型多形性,表面积、最大直径、周长、等效圆直径和形状因子,并作比较分析。四种细菌Ｌ型及Ｌ型中的巨形体在上述５项形态学参数指标上除形状因子外均较细菌型明显增大,而形状因子参数明显小于细菌型。细菌Ｌ型之间在形态学指标上的差异有显著性（ｐ＜０．０５）。形态定量分析能准确客观地反映形态定性观察的结果以及细菌Ｌ型的形态变异程度,为细菌Ｌ型的形态定量分析提供依据。     

A protein–protein interaction map reveals that the Coxiella burnetii effector CirB inhibits host proteasome activity

Coxiella burnetii is the etiological agent of the zoonotic disease Q fever, which is featured by its ability to replicate in acid vacuoles resembling the lysosomal network. One key virulence determinant of C. burnetii is the Dot/Icm system that transfers more than 150 effector proteins into host cells. These effectors function to construct the lysosome-like compartment permissive for bacterial replication, but the functions of most of these effectors remain elusive. In this study, we used an affinity tag purification mass spectrometry (AP-MS) approach to generate a C. burnetii-human protein-protein interaction (PPI) map involving 53 C. burnetii effectors and 3480 host proteins. This PPI map revealed that the C. burnetii effector CBU0425 (designated CirB) interacts with most subunits of the 20S core proteasome. We found that ectopically expressed CirB inhibits hydrolytic activity of the proteasome. In addition, overexpression of CirB in C. burnetii caused dramatic inhibition of proteasome activity in host cells, while knocking down CirB expression alleviated such inhibitory effects. Moreover, we showed that a region of CirB that spans residues 91–120 binds to the proteasome subunit PSMB5 (beta 5). Finally, PSMB5 knockdown promotes C. burnetii virulence, highlighting the importance of proteasome activity modulation during the course of C. burnetii infection.     

三江源牧户参与草地生态保护的意愿

牧民对环境保护的响应是生态保护战略有效实施的关键,主体参与意愿直接影响生态保护项目实施的成效和可持续性,是区域生态经济可持续发展的前提。通过对三江源地区约283户藏族牧民通过翻译进行结构式访谈,采用Logistic模型从主体角度探讨了牧户愿意参与生态保护行为响应的主要影响因素。结果表明:(1)三江源区域约87%的牧户认为生态保护对牧户有好处,但受各种内外部因素的影响只有近70%的牧户是在政府主导下基于有限理性而被动的参与生态保护行为响应。(2)三江源牧户参与生态保护行为响应的意愿主要受当地政府的保护力度及牧户对生态保护外部性的认知水平、生计水平、外界接触程度、工作机会的正影响,并受牧户的年龄、离中心城镇的距离和区域气候恶劣情况等因素的负影响,系数依次为:2.22、3.98、1.93、2.26、1.48、-1.63、-2.43、-0.92。(3)牧户的生计水平、退化感知和外部性认知是影响三江源牧户参与生态保护意愿的关键因素,牧户参与生态保护意愿的概率不仅仅是牧户出于自身利益和未知风险考虑下被动的响应,更是当地政府的环境知识宣传和保护投入影响下个体的抉择结果。(4)创造更多的就业机会和解决牧户的单一化生计问题,构建完善地生态补偿机制让牧户分享生态保护的外部性效益,并激励牧户主动参与生态保护行为响应,才能最终实现区域生态保护、牧户幸福和生态经济可持续发展的多赢局面。     

一株PCBs降解菌的降解特性及发酵条件优化

【目的】针对一株多氯联苯的高效降解菌,考察其对多氯联苯(PCBs)的降解特性,并对降解条件进行优化。【方法】以不同浓度的2,4,4′-TCB与3,3′,4,4′-TCB为唯一碳源,研究苜蓿中华根瘤菌(Sinorhizobium melilon)对不同多氯联苯的降解转化能力,并进行发酵条件优化以及共代谢试验。【结果】接入菌株转化7 d后,随着底物浓度的增加,该菌对2,4,4′-TCB的降解能力呈下降趋势。在最低浓度1.0 mg/L时降解率最高,为93.3%;而在最高浓度50.0 mg/L时为65.1%。对于较难降解的四氯联苯3,3′,4,4′-TCB,菌株在最低浓度1.0 mg/L时降解率为56.2%,最高浓度25.0 mg/L时为22.8%。在温度30°C、pH 7.0、接种量4.5 mL、装液量25 mL时,获得菌株转化10.0 mg/L 2,4,4′-TCB的最优发酵条件,7 d的降解率由原来的54.8%提高到83.6%。柠檬烯、香芹酮及甘露醇作为共代谢底物也可较好地提高菌株降解效果。【结论】苜蓿中华根瘤菌对PCBs有很好的降解效果,研究结果对PCBs的微生物降解及环境中PCBs的生物修复具有较好的意义和应用价值。     

我国苏云金杆菌研究60年

综述了我国近60年来的苏云金杆菌研究进展,包括资源收集及分类鉴定、Bt新基因的发掘及组学研究、病理学与作用机制、毒力测定、产品标准化、产业化,并就Bt生物农药存在的问题及改善途径进行了探讨。     

BB染色体组野生种花生LTR反转录转座子RT基因的多样性分析

利用简并PCR技术从野生花生种(Arachis ipaensis Krapov. et W. C. Greg.)的基因组中扩增分离Ty1-copia类(1类)和Ty3-gypsy类(2类)反转录转座子RT基因,并对其序列特征、多样性、系统进化关系及转录活性进行分析。结果显示:对于1和2类RT基因,目的条带分别约为260和430 bp;分离获得了23和32条序列,长度范围分别为262～266、395～435 bp;AT所占比例分别为61.60%～69.17%和55.79%～61.34%;核苷酸序列间相似性分别为52.5%～98.9%和45.0%～98.8%,氨基酸序列间相似性分别为39.8%～100%和9.0%～97.2%。其中2类基因的异质性高于1类;1类和2类基因分别有3条和15条发生了无义突变,2类的无义突变发生率远高于1类。1类基因的保守基序保守性较高,2类的保守基序呈一定程度的变异。代表序列的蛋白质三级结构基本类似。聚类分析结果显示,1类和2类基因可被分为5个和6个家族。1类和2类基因都有部分序列与其他物种的RT基因序列亲缘关系较近,表明它们之间可能存在两类反转录转座子的横向传...     

Toxoplasma infection in male mice alters dopamine-sensitive behaviors and host gene expression patterns associated with neuropsychiatric disease

During chronic infection, the single celled parasite, Toxoplasma gondii, can migrate to the brain where it has been associated with altered dopamine function and the capacity to modulate host behavior, increasing risk of neurocognitive disorders. Here we explore alterations in dopamine-related behavior in a new mouse model based on stimulant (cocaine)-induced hyperactivity. In combination with cocaine, infection resulted in heightened sensorimotor deficits and impairment in prepulse inhibition response, which are commonly disrupted in neuropsychiatric conditions. To identify molecular pathways in the brain affected by chronic T. gondii infection, we investigated patterns of gene expression. As expected, infection was associated with an enrichment of genes associated with general immune response pathways, that otherwise limits statistical power to identify more informative pathways. To overcome this limitation and focus on pathways of neurological relevance, we developed a novel context enrichment approach that relies on a customized ontology. Applying this approach, we identified genes that exhibited unexpected patterns of expression arising from the combination of cocaine exposure and infection. These include sets of genes which exhibited dampened response to cocaine in infected mice, suggesting a possible mechanism for some observed behaviors and a neuroprotective effect that may be advantageous to parasite persistence. This model offers a powerful new approach to dissect the molecular pathways by which T. gondii infection contributes to neurocognitive disorders.     

LRP6 downregulation promotes cardiomyocyte proliferation and heart regeneration

The adult mammalian heart is thought to be a terminally differentiated organ given the postmitotic nature of cardiomyocytes. Consequently, the potential for cardiac repair through cardiomyocyte proliferation is extremely limited. Low-density lipoprotein receptor-related protein 6 (LRP6) is a Wnt co-receptor that is required for embryonic heart development. In this study we investigated the role of LRP6 in heart repair through regulation of cardiomyocyte proliferation. Lrp6 deficiency increased cardiomyocyte cell cycle activity in neonatal, juvenile and adult mice. Cardiomyocyte-specific deletion of Lrp6 in the mouse heart induced a robust regenerative response after myocardial infarction (MI), led to reduced MI area and improvement in left ventricular systolic function. In vivo genetic lineage tracing revealed that the newly formed cardiomyocytes in Lrp6-deficient mouse hearts after MI were mainly derived from resident cardiomyocytes. Furthermore, we found that the pro-proliferative effect of Lrp6 deficiency was mediated by the ING5/P21 signaling pathway. Gene therapy using the adeno-associated virus (AAV)9 miRNAi-Lrp6 construct promoted the repair of heart injury in mice. Lrp6 deficiency also induced the proliferation of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Our study identifies LRP6 as a critical regulator of cardiomyocyte proliferation, which may lead to the development of a novel molecular strategy to promote myocardial regeneration and repair.Subject terms: Cell-cycle exit, Cytokinesis