肺细胞色素氧化酶体视学测量法在高压氧研究中的应用

采用超微结构细胞化学方法,用计算机图象分析仪测量计算了小鼠肺毛细血管和Ⅱ型肺泡线粒体、线粒体膜和嵴上细胞色素氧化酶活性变化的二维形态学、三维立体学定量参数。结果表明,毛细血管线粒体面积较Ⅱ型肺泡细胞小,但体密度却大;毛细血管线粒体细胞色素氧化酶阳性反应面积比正型肺泡细胞小,但酶反应体密度却大、暴露于高压氧（０．５ＭＰａ）后小鼠Ⅱ型肺泡细胞线粒体细胞色素氧化酶活性降低。     

Anti-breast cancer sinomenine derivatives via mechanisms of apoptosis induction and metastasis reduction

Sinomenine, a morphinane-type isoquinoline-derived alkaloid, was first isolated from stems and roots of Sinomenium diversifolius (Miq.) in 1920. Later discovery by researchers confirmed various essential biological efficacy sinomenine exerted in vitro and in vivo. In this study, a series of 15 sinomenine/furoxan hybrid compounds were designed and synthesised in search of a TNBC drug candidate. Some of the target compounds exhibited strong antiproliferative activities against cancer cell lines, especially for TNBC cells, compared to positive controls. Among them, hybrid 7Cc exerted superior cytotoxic effects on cancer cell lines with exceptionally low IC₅₀ (0.82 μM) against MDA-MB-231 cells with the highest safety index score. Further studies in mechanism displayed that 7Cc could induce an S phase cell cycle arrest, stimulate apoptosis in MDA-MB-231 cells, disrupt mitochondrial membrane potential and exert a genotoxic effect on DNA in cancer cells. In addition, 7Cc also notably inhibited MDA-MB-231 cells in both migration, invasion and adhesion.     

嗜麦芽窄食单胞菌OUC_Est10全基因组测序及脂类水解酶多样性分析

[目的]本研究的目的是研究嗜麦芽窄食单胞菌OUC_Est10中脂类水解酶的多样性。[方法]使用离子交换层析、全基因组测序和异源表达三种方法研究嗜麦芽窄食单胞菌OUC_Est10中脂类水解酶的多样性。[结果]离子交换层析结果显示嗜麦芽窄食单胞菌OUC_Est10可以分泌多种脂类水解酶。通过全基因组测序,我们给出了该菌的全基因组序列,该基因组大小为4668743 bp,GC含量为66.25%。通过详细的基因组序列分析,我们从该基因组中找到33个可能具有脂类水解酶活性的假定基因。通过异源表达OUC_Est10中的5个假定脂类水解酶基因,来研究其催化特性的多样性,结果显示这些脂类水解酶具有不同的催化特性。[结论]我们证明了嗜麦芽窄食单胞菌OUC_Est10拥有多样的脂类水解酶,这暗示了它在不同领域中的应用潜力。     

克雷伯氏菌属(Klebsiella)细菌比较基因组学分析揭示多复制子抗性质粒介导广泛的抗性基因传播

[目的] 研究克雷伯氏菌与多复制子抗性质粒间的关系,分析细菌携带多复制子质粒对抗生素环境的响应机制。[方法] 以2018-2020年分离的56株不同来源克雷伯氏菌（Klebsiella sp.）分离株为研究对象,利用微量肉汤稀释法评估其多重耐药表型,对分离菌株进行全基因组测序（WGS）,通过细菌全基因组关联分析（BGWAS）技术和比较基因组学方法深入解析多复制子抗性质粒形成的机制。[结果] 耐药表型分析发现野生动物来源的菌株具有更广的耐药谱系,总体Klebsiella sp.对氨苄西林表现出很高的耐药率（80.36%）,尤其是马来穿山甲来源菌株对头孢类抗生素高度耐受,同时对氯霉素、左氧氟沙星和复方新诺明等药物耐受,基因组分析发现这些菌株携带了抗性质粒和更多的抗生素抗性基因。进一步对69个质粒序列分析,发现有28个质粒为多复制子质粒,主要携带bla_CTX-M-15、bla_CTX-M-14、bla_CTX-M-55、bla_OXA-1和bla_TEM-1等β-内酰胺酶基因。细菌携带质粒类型分析认为Klebsiella pneumoniae可能是多复制子质粒的重要宿主,质粒骨架与结构分析发现多复制子质粒多由2个或2个以上单个质粒融合而成,携带此类质粒的菌株不仅获得了更广的耐药表型,而且在全球传播扩散分布逐年增加,因此产生对抗生素环境更强的适应性。[结论] 多重耐药性细菌呈现的表型与携带的多复制子质粒有关,相比较下多复制子质粒比非多复制子质粒有更强的抗性基因携带能力,或许是细菌在强大的抗生素压力下产生的重要响应机制。本研究对于未来探索细菌抗性基因的传播扩散机制具有重要意义。     

传统生态知识的民族生态学分析框架

传统生态知识是民族生态学研究的核心范畴,国外已有多年的研究基础,国内的相关研究正是方兴未艾。通过文献查询和比较分析等方法,介绍了国外已有的传统生态知识的民族生态学分析框架的3个模型:知识-实践-信仰的三角形框架;本土经验知识-资源管理知识-社会制度知识-世界观知识的四椭圆框架;事实观察-管理体系-旧有及当下利用-伦理价值-文化特征-宇宙观的五边形框架。结合田野调查研究,提出了一个立体的传统生态知识的民族生态学分析模型,强调了传统生态知识的动态特征,将民族对于自然的被动认识和主动认知、民族人际关系规范、民族的哲学与伦理等方面所呈现出的传统生态知识进行了分类,从而为民族生态学的调查研究提供了方法上的建议。最后总结了这个立体的传统生态知识分析框架的意义,并建议在我国的生态文明建设中发展和应用民族生态学。     

Transcriptomic Profiling of Spleen in Grass-Fed and Grain-Fed Angus Cattle

The grass-fed cattle obtain nutrients directly from pastures containing limited assimilable energy but abundant amount of fiber; by contrast, grain-fed steers receive a diet that is comprised mainly of grains and serves as an efficient source of high-digestible energy. Besides energy, these two types of diet differ in a large number of nutritional components. Additionally, animals maintained on rich-energy regimen are more likely to develop metabolic disorders and infectious diseases than pasture raised individuals. Thus, we hypothesize that spleen–a relevant immune organ–may function differently under disparate regimes. The objective of this study was to find the differentially expressed genes in the spleen of grass-fed and grain-fed steers, and furtherly explore the potential involved biopathways. Through RNA sequencing (RNA-Seq), we detected 123 differentially expressed genes. Based on these genes, we performed an Ingenuity Pathway Analysis (IPA) and identified 9 significant molecular networks and 13 enriched biological pathways. Two of the pathways, Nur77 signaling in T lymphocytes and calcium-induced T lymphocyte apoptosis which are immune related, contain a pair of genes HLA-DRA and NR4A1 with dramatically altered expression level. Collectively, our results provided valuable insights into understanding the molecular mechanism of spleen under varied feeding regimens.     

Identification of noncytotoxic and IL-10-producing CD8+AT2R+ T cell population in response to ischemic heart injury

Emerging evidence suggests a cardioprotective role of the angiotensin AT2R, albeit the underlying cellular mechanisms are not well understood. We aimed in this article to elucidate a potential role of cardiac angiotensin AT2R in regulating cellular immune response to ischemic heart injury. Seven days after myocardial infarction in rats, double-immunofluorescence staining showed that AT2R was detected in a fraction of CD8(+) T cells infiltrating in the peri-infarct myocardium. We developed a method that allowed the isolation of myocardial infiltrating CD8(+)AT2R(+) T cells using modified MACS, and further characterization and purification with flow cytometry. Although the CD8(+)AT2R(-) T cells exhibited potent cytotoxicity to both adult and fetal cardiomyocytes (CMs), the CD8(+)AT2R(+) T cells were noncytotoxic to these CMs. The CD8(+)AT2R(+) T cells were characterized by upregulated IL-10 and downregulated IL-2 and INF-γ expression when compared with CD8(+)AT2R(-) T cells. We further showed that IL-10 gene expression was enhanced in CD8(+) T cells on in vitro AT2R stimulation. Importantly, in vivo AT2R activation engendered an increment of CD8(+)AT2R(+) T cells and IL-10 production in the ischemic myocardium. In addition, intramyocardial transplantation of CD8(+)AT2R(+) T cells (versus CD8(+)AT2R(-)) led to reduced ischemic heart injury. Moreover, the CD8(+)AT2R(+) T cell population was also demonstrated in human peripheral blood. Thus, we have defined the cardioprotective CD8(+)AT2R(+) T cell population, which increases during ischemic heart injury and contributes to maintaining CM viability and providing IL-10, hence revealing an AT2R-mediated cellular mechanism in modulating adaptive immune response in the heart.     

絮凝活性产紫青霉EL-02的分离鉴定及其絮凝活性初探

【目的】分离鉴定有絮凝活性真菌,同时对其絮凝活性进行初步研究。【方法】采用梯度稀释、平板划线、18SrDNA检测等方法分离鉴定絮凝活性菌株。通过高速离心、超声破碎、乙醇沉淀、定性试验等方法确定絮凝活性物质性质。【结果】从渤海湾海岸土壤样品中分离筛选出一株有较高絮凝活性的真菌,经鉴定为产紫青霉(Penicillium purpurogenum),命名为产紫青霉EL-02(P.purpurogenum EL-02)。超声破碎试验证实其絮凝活性主要存在于发酵上清液。根据絮凝活性曲线,确定4d为积累絮凝活性产物的最佳发酵时间。乙醇沉淀法获得该菌株絮凝活性物质。经鉴定该菌株所产絮凝活性物质为糖类,且其活性在pH2-11,温度-70℃-100℃范围内保持稳定。【结论】分离筛选到一株有絮凝活性的产紫青霉EL-02,经鉴定其产生糖类絮凝活性物质。     

高频喷射通气治疗海水淹溺肺水肿时兔肺钙离子超微结构变化

用海水灌注的方法诱导海水淹溺肺水肿（ＰＥ－ＳＷＤ）, 采用高频喷射通气（ＨＦＪＶ） 治疗, 应用血气酸碱分析仪对兔动脉血气酸碱三项指标进行自动检测, 并采用Ｃａ２＋ 超微结构方法观察肺内Ｃａ２＋ 变化。结果表明肺细胞内Ｃａ２＋ 超负荷与ＰＥ－ＳＷＤ发生有关, ＨＦＪＶ 对改善ＰＥ－ＳＷＤ症状有明显作用, 同时, 细胞内Ｃａ２＋ 超负荷状况明显缓解