携带IL-2/NK4双基因减毒沙门氏菌TPIN的遗传稳定性研究

对携带IL-2/NK4双基因减毒沙门氏菌TPIN的遗传稳定性进行研究。将TPIN连续传40代,取10代、20代、30代、40代菌落进行鉴定,包括菌落和菌体的形态、质粒的纯度和浓度、目的基因片段、双酶切结果及转染人肝癌细胞(HepG2)后目的基因体外表达活性。结果表明,每10代的菌落和菌体形态一致,其所含质粒的纯度和浓度在传代过程中无明显变化,PCR扩增的目的基因条带大小一致,双酶切鉴定结果正确,转染HepG2细胞后,每代间所表达目的蛋白浓度无明显差异。TPIN作为消化道肿瘤的生物治疗药物,在遗传方面有较好的稳定性,可进一步进行大量生产。     

C6-Ceramide Nanoliposomes Target the Warburg Effect in Chronic Lymphocytic Leukemia

Ceramide is a sphingolipid metabolite that induces cancer cell death. When C6-ceramide is encapsulated in a nanoliposome bilayer formulation, cell death is selectively induced in tumor models. However, the mechanism underlying this selectivity is unknown. As most tumors exhibit a preferential switch to glycolysis, as described in the “Warburg effect”, we hypothesize that ceramide nanoliposomes selectively target this glycolytic pathway in cancer. We utilize chronic lymphocytic leukemia (CLL) as a cancer model, which has an increased dependency on glycolysis. In CLL cells, we demonstrate that C6-ceramide nanoliposomes, but not control nanoliposomes, induce caspase 3/7-independent necrotic cell death. Nanoliposomal ceramide inhibits both the RNA and protein expression of GAPDH, an enzyme in the glycolytic pathway, which is overexpressed in CLL. To confirm that ceramide targets GAPDH, we demonstrate that downregulation of GAPDH potentiates the decrease in ATP after ceramide treatment and exogenous pyruvate treatment as well as GAPDH overexpression partially rescues ceramide-induced necrosis. Finally, an in vivo murine model of CLL shows that nanoliposomal C6-ceramide treatment elicits tumor regression, concomitant with GAPDH downregulation. We conclude that selective inhibition of the glycolytic pathway in CLL cells with nanoliposomal C6-ceramide could potentially be an effective therapy for leukemia by targeting the Warburg effect.     

The Relationship between RTS,S Vaccine-Induced Antibodies,CD4+ T Cell Responses and Protection against Plasmodium falciparum Infection

Vaccination with the pre-erythrocytic malaria vaccine RTS,S induces high levels of antibodies and CD4⁺ T cells specific for the circumsporozoite protein (CSP). Using a biologically-motivated mathematical model of sporozoite infection fitted to data from malaria-naive adults vaccinated with RTS,S and subjected to experimental P. falciparum challenge, we characterised the relationship between antibodies, CD4⁺ T cell responses and protection from infection. Both anti-CSP antibody titres and CSP-specific CD4⁺ T cells were identified as immunological surrogates of protection, with RTS,S induced anti-CSP antibodies estimated to prevent 32% (95% confidence interval (CI) 24%–41%) of infections. The addition of RTS,S-induced CSP-specific CD4⁺ T cells was estimated to increase vaccine efficacy against infection to 40% (95% CI, 34%–48%). This protective efficacy is estimated to result from a 96.1% (95% CI, 93.4%–97.8%) reduction in the liver-to-blood parasite inoculum, indicating that in volunteers who developed P. falciparum infection, a small number of parasites (often the progeny of a single surviving sporozoite) are responsible for breakthrough blood-stage infections.     

慢性心力衰竭患者血清Galectin-3、hs-cTnT、Cys C和PTX-3水平变化及临床意义

目的:研究慢性心力衰竭(CHF)患者血清半乳糖凝聚素-3(Galectin-3)、高敏肌钙蛋白-T(hs-cTnT)、胱抑素C(Cys C)和正五聚体蛋白-3(PTX-3)水平变化及临床意义。方法:选择2017年2月～2018年6月我院收治的CHF患者100例,按照美国心脏病协会(NYHA)心功能分级标准将其分成NYHAⅡ级组39例、NYHAⅢ级组33例、NYHAⅣ级组28例,根据随访1年患者预后情况将主要心脏不良事件患者记作预后不良组(n=27),其余记为预后优良组(n=73),另取同期于我院进行体检的健康者30例作为对照组。分别比较CHF患者和对照组的心功能指标、血清Galectin-3、hs-cTnT、Cys C、PTX-3水平,分析CHF患者上述指标的相关性及其与CHF预后情况的关系。结果:对照组、NYHAⅡ级组、NYHAⅢ级组、NYHAⅣ级组左心室舒张末期内径(LVEDD)呈逐渐增大趋势,而左心室射血分数(LVEF)呈逐渐降低趋势(P0.05)。对照组、NYHAⅡ级组、NYHAⅢ级组、NYHAⅣ级组血清Galectin-3、hs-cTnT、Cys C和PTX-3水平呈逐渐升高趋势(P0.05)。经Pearson相关性分析可得:CHF患者LVEDD与血清Galectin-3、hs-cTnT、Cys C、PTX-3水平呈正相关关系,而LVEF与血清Galectin-3、hs-cTnT、Cys C、PTX-3水平呈负相关关系(P0.05)。CHF预后优良组血清Galectin-3、hs-cTnT、Cys C、PTX-3水平均低于预后不良组(P0.05)。结论:CHF血清Galectin-3、hs-cTnT、Cys C和PTX-3水平均呈明显高表达,且与患者的病情严重程度呈密切相关,临床上可通过检测上述指标水平,继而为CHF临床诊断、预后评估提供参考依据。     

Allogeneic HLA-A*02-restricted WT1-specific T cells from mismatched donors are highly reactive but show off-target promiscuity

T cells recognizing tumor-associated Ags such as Wilms tumor protein (WT1) are thought to exert potent antitumor reactivity. However, no consistent high-avidity T cell responses have been demonstrated in vaccination studies with WT1 as target in cancer immunotherapy. The aim of this study was to investigate the possible role of negative thymic selection on the avidity and specificity of T cells directed against self-antigens. T cell clones directed against the HLA-A*0201-binding WT1(126-134) peptide were generated from both HLA-A*02-positive (self-HLA-restricted) and HLA-A*02-negative [nonself (allogeneic) HLA [allo-HLA]-restricted] individuals by direct ex vivo isolation using tetramers or after in vitro priming and selection. The functional avidity and specificity of these T cell clones was analyzed in-depth. Self-HLA-restricted WT1-specific clones only recognized WT1(126-134) with low avidities. In contrast, allo-HLA-restricted WT1 clones exhibited profound functional reactivity against a multitude of HLA-A*02-positive targets, even in the absence of exogenously loaded WT1 peptide, indicative of Ag-binding promiscuity. To characterize this potential promiscuity, reactivity of the T cell clones against 400 randomly selected HLA-A*0201-binding peptides was investigated. The self-HLA-restricted WT1-specific T cell clones only recognized the WT1 peptide. In contrast, the allo-HLA-restricted WT1-reactive clones recognized besides WT1 various other HLA-A*0201-binding peptides. In conclusion, allogeneic HLA-A*02-restricted WT1-specific T cells isolated from mismatched donors may be more tumor-reactive than their autologous counterparts but can show specific off-target promiscuity of potential clinical importance. As a result of this, administration of WT1-specific T cells generated from HLA-mismatched donors should be performed with appropriate precautions against potential off-target effects.     

喀斯特断陷盆地区不同恢复阶段群落物种组成与多样性特征

研究自然植被恢复过程中的物种组成、群落结构及生物多样性的变化,能够为人工促进植被恢复的树种选择与群落结构的优化配置提供重要依据。本研究以空间代替时间对喀斯特断陷盆地典型区云南省建水县不同天然植被(草丛、灌丛、乔木林)进行群落学调查,对不同恢复阶段的植物群落按乔木、灌木、草本进行分层,分析各恢复阶段植物群落的物种组成、水平和垂直结构、生物多样性。结果表明:在总面积为3200 m²的12个样地中,共记录43科72属94种维管束植物,优势种以壳斗科(Fagaceae)、鼠李科(Rhamnaceae)、紫金牛科(Myrsinaceae)、蔷薇科(Rosaceae)、木犀科(Oleaceae)等科的植物为主;在草丛→灌丛→乔木林的恢复过程中,群落物种组成中的科数、属数、种数逐渐增加,低矮和小径级植物个体数所占比例逐渐减少,但整体仍以低矮的小径级植物为主。草本植物的丰富度和Shannon指数在植被恢复的初期即草丛阶段最大,而均匀度指数则以灌丛阶段最大;木本植物的丰富度和Shannon指数随着植被的恢复逐渐增大,但均匀度指数随着植被的恢复逐渐下降;随着植被的恢复,草本层和乔木层...     

红松人工林优势木竞争指数影响因子

在温带湿润气候区东段不同立地条件下的红松人工林内设置面积为600 m~2(20 m×30 m)的70块矩形固定样地。在每个样地内,选取5颗最高且长势较好的优势木作为研究的对象木,用Voronoi图确定优势木相应的竞争木,测定每一块样地内优势木与竞争木之间的距离。采用Hegyi单木竞争指数模型,分析优势木在不同样地水平上的种内竞争强度,探讨林分生长因子、地形因子、土壤养分因子对优势木竞争指数的影响,并对这3类因子与优势木竞争指数进行相应的拟合和相关性分析。结果表明：红松人工林优势木竞争强度随着优势木胸径的增大而变小,并且两者之间的关系服从幂函数;红松优势木的竞争指数与其树高、胸径、冠幅呈极显著的相关关系(P<0.01);坡向、坡位、海拔对竞争指数影响极显著(P<0.01);红松人工林优势木竞争指数的大小与土壤氮磷钾含量均呈极显著的相关关系(P<0.01);pH值对优势木竞争指数的影响不显著。当红松人工林优势木平均胸径达到45cm,优势木平均树高和冠幅大于周围竞争木时,其对周围资源的利用程度增大,林木会发生自然稀疏现象,其所受的竞争压力减小。红松喜光性强,对水分的要求高,...     

海拔高度对2型糖尿病患者循环内皮祖细胞及低氧诱导因子的影响*

目的: 探讨不同海拔地区2型糖尿病患者循环内皮祖细胞数量及相关检测指标的变化情况,为2型糖尿病血管并发症的研究和治疗提供依据。方法: 选取386 m低海拔地区(咸阳市)和1 520 m高海拔地区(兰州市)各一家医院内被诊断的2型糖尿病患者(25人/29人)和健康体检者(20人/20人)。用全自动生化分析仪检测两组人群的血脂、血糖和糖化血红蛋白指标,用酶联免疫吸附试验(ELISA)检测低氧诱导因子-1α(HIF-1α)的浓度,用流式细胞仪测定外周血循环内皮祖细胞(EPCs)的数量。结果: 无论在低海拔还是高海拔地区,糖尿病组较健康组循环EPCs数量降低(P＜0.01),体质指数(BMI)、腰臀比(WHR)、甘油三酯(TG)、空腹血糖(FBG)及糖化血红蛋白含量(HbAlc)增高(P＜0.05);与低海拔组相比,无论高海拔的糖尿病患者还是健康者,HIF-1α的表达水平均明显增加(P＜0.05),而循环EPCs数量明显降低(P＜0.05),且有循环EPCs数量健康者高于2型糖尿病无血管并发症者高于2型糖尿病伴血管并发症者的表现(P＜0.05)。结论: 海拔高度增加,2型糖尿病(T2DM)患者体内HIF-1α表达水平增加,循环EPCs数量降低,且与其血管病变程度密切相关。因此有望通过对高海拔地区T2DM患者进行EPCs的移植来实现对糖尿病血管并发症的预防和改善。     

微塑料对湖泊生态系统初级生产者的影响

塑料的广泛应用导致大量微塑料进入环境,尤其是水环境,从而产生环境风险。浮游植物等自养生物是湖泊系统的主要初级生产者,是湖泊食物链的关键组成部分,为食物链的上游提供能量和物质基础。同时,浮游植物也是对微塑料响应最敏感的类群。了解湖泊浮游植物等初级生产者对微塑料的响应是探究微塑料对湖泊生态系统功能影响的重要基础。总结了全球湖泊生态系统微塑料的丰度、类型、尺寸、来源等分布特征,系统分析了微塑料暴露对浮游植物等初级生产者细胞结构、基因表达和生长,以及对浮游植物叶绿素a含量、光合活性的影响,并总结了其中的影响机制。总体而言,微塑料会降低初级生产者的叶绿素a含量,剂量越高、尺寸越小,这种抑制作用越强烈;同时,微塑料也会作用于初级生产者,造成细胞膜损伤、DNA损伤,调控其相关功能基因表达,抑制其生长和光合活性等。然而,湖泊生态系统微塑料的实际检出浓度远低于室内暴露实验中的添加剂量,微塑料结构和组分也更为复杂,野外观测结果与室内培养实验之间还不能建立直接的对应关系。因此,未来的相关研究应集中在如何有效联系野外观测结果与室内培养实验结果,进一步聚焦建立可靠的、可应用推广的微塑料浓度与初级生产者之间的剂量-效应关系模型,探究微塑料对湖泊初级生产者的作用机制,为刻画微塑料对湖泊生态系统初级生产者及其功能的作用机制提供科学支撑。