用于高灵敏可视化检测松材线虫的闭管等温扩增法

建立了一种基于环介导等温核酸扩增技术(LAMP)的松材线虫高灵敏可视化闭管检测方法。针对松材线虫核糖体DNA的序列保守区域设计LAMP引物,通过优化LAMP体系中的Mg2+、甜菜碱浓度和反应温度等因素,建立了环介导等温扩增法;并结合蜡封反应管对产物进行检测,检测结果可直接通过肉眼观察SYBR Green I荧光显色进行判定。结果表明,本方法可检测到低至10拷贝/管的松材线虫核酸片段,可对单条线虫进行检测,并且具有很高的特异性,能区分检测松材线虫与拟松材线虫。由于整个反应恒温进行,无需热循环仪;闭管检测极大地降低了扩增产物交叉污染的风险;检测速度快,整个检测过程只需40 min,为松材线虫的现场快速筛检提供了一种简便、高灵敏、高特异的工具。     

Vitamin A Supplementation in Early Life Enhances the Intestinal Immune Response of Rats with Gestational Vitamin A Deficiency by Increasing the Number of Immune Cells

Vitamin A is a critical micronutrient for regulating immunity in many organisms. Our previous study demonstrated that gestational or early-life vitamin A deficiency decreases the number of immune cells in offspring. The present study aims to test whether vitamin A supplementation can restore lymphocyte pools in vitamin A-deficient rats and thereby improve the function of their intestinal mucosa; furthermore, the study aimed to identify the best time frame for vitamin A supplementation. Vitamin A-deficient pregnant rats or their offspring were administered a low-dose of vitamin A daily for 7 days starting on gestational day 14 or postnatal day 1, day 14 or day 28. Serum retinol concentrations increased significantly in all four groups that received vitamin A supplementation, as determined by high-performance liquid chromatography. The intestinal levels of secretory immunoglobulin A and polymeric immunoglobulin receptor increased significantly with lipopolysaccharide challenge in the rats that received vitamin A supplementation starting on postnatal day 1. The rats in this group had higher numbers of CD8⁺ intestinal intraepithelial lymphocytes, CD11_C ⁺ dendritic cells in the Peyer''s patches and CD4⁺CD25⁺ T cells in the spleen compared with the vitamin A-deficient rats; flow cytometric analysis also demonstrated that vitamin A supplementation decreased the number of B cells in the mesenteric lymph nodes. Additionally, vitamin A supplementation during late gestation increased the numbers of CD8⁺ intestinal intraepithelial lymphocytes and decreased the numbers of B lymphocytes in the mesenteric lymph nodes. However, no significant differences in lymphocyte levels were found between the rats in the other two vitamin A supplement groups and the vitamin A-deficient group. In conclusion, the best recovery of a subset of lymphocytes in the offspring of gestational vitamin A-deficient rats and the greatest improvement in the intestinal mucosal immune response are achieved when vitamin A supplementation occurs during the early postnatal period.     

中国狭长蝽属初记(半翅目:长蝽科:杆长蝽亚科)

狭长蝽属(Dimorphopterus Stal)为杆长蝽亚科(Blissinae)中较大的一属。身体常成两侧平行的小杆状,多数尚较粗壮,亦有甚狭长者,体黑,被平伏或半直立毛,翅革片白色而有深色斑,常出现短翅型个体。多生活于禾本科植物上,北方地区为害高粱的“高粱长蝽”即属此类。 本属的概念和范围几经变动:Stal 1872年建立此属时,用身体较狭,触角短于头、胸     

光照和氮磷供应比对木荷生长及化学计量特征的影响

光照和养分限制是影响林下植物生长和更新的关键影响因素,以亚热带主要常绿树种木荷（Schima superba）实生幼苗为试验对象,研究了不同光照（全光照、遮阴即45%全光照）和N、P供应比例（5,15,45）对幼苗生长和化学计量特征的影响。结果表明：（1）遮阴不仅严重抑制了木荷各器官和单株生物量积累,更加剧了P限制。尽管N、P添加对木荷生长没有显著促进作用,但N、P供应比例为5时的性状组合更有利于木荷后期生长,但高N、P供应比例可能导致P限制。（2）遮阴下叶N、P含量显著增加,但叶C/N和C/P比显著降低;不同光照处理组中各器官及总N含量均随N、P供应比例增大而显著增加,而C/N比逐渐降低;P的分配格局发生改变,全光照组各器官P含量为茎 > 叶 > 根,遮阴组各器官P含量为根 > 茎 > 叶。（3）随N、P供应比例增加或光照强度降低,木荷均趋向降低根冠比和根质比、增加叶质比或茎质比。（4）木荷生物量与各器官N、P含量、叶质比呈极显著负相关,而与C/N和C/P比及根冠比、茎质比、根质比呈极显著正相关。光强和N、P比例变化均显著影响了木荷幼苗的养分利用特征,因而木荷作为伴生树种优化林分环境对其早期生长具有重要意义。     

SIRT1与肝细胞癌

肝细胞癌(hepatocellular carcinoma, HCC)是世界上最常见的癌症之一.然而,就目前现状而言,HCC的治疗效果还很有限.作为一个依赖于烟酰胺腺嘌呤二核苷酸(NAD+)的去乙酰化酶, SIRT1(silent mating type information regulation 2 homolog 1 )参与了代谢、应激反应、衰老以及肿瘤的演进等许多重要的生物学进程.临床研究显示,SIRT1在HCC患者中异常高表达,并可预测其不良预后;进一步的研究表明,SIRT1在HCC演进中发挥了关键作用,且作用范围广泛,分子机制复杂.这提示,SIRT1有望成为新的HCC治疗靶点和诊断、预后标志物.本文拟对SIRT1在HCC的演进和预后中的具体作用及其潜在分子机制作一总结,并就SIRT1作为肝癌治疗靶点和诊断、预后标志物的可行性做出探讨.     

糖尿病肾病发病机制研究进展

糖尿病肾病(DN)是糖尿病(DM)最常见的慢性并发症,也是终末期肾病(ESRD)的主要原因,其治疗费用巨大。其发病机制主要涉及遗传易感性、糖代谢紊乱、肾血流动力学的改变、细胞因子、炎症机制已及氧化应激等方面。本文就以上作用机制的最新研究进展作一综述。     

芽胞抗力快速弱化技术研究

以L-丙氨酸缓冲液为发芽剂,结合芬顿反应原理,观察发芽-氧化损伤效应对芽胞的杀灭效果,以期为新型炭疽疫源地净化方法的深入研究奠定基础。以腊样芽胞为试验菌,采用透射电镜、激光扫描共聚焦显微镜、活菌计数等方法观察芽胞发芽过程的超微结构、核酸含量变化,以及在芬顿反应的联合作用下发芽体的活性变化。在20~30 min的发芽过程中,芽胞核心密度降低,核心与皮质、皮质与外壁之间界限模糊,芽胞外壁和芽胞衣有破裂,通透性增加,进一步有皮质消失、细胞核与细胞质融合、细胞膜基本形成的现象;发芽体荧光强度不断增加,显示菌体中核酸的活性和含量不断增加;发芽体对化学因子的抗力明显下降,H2O2浓度为0.20 mol/L的Fenton反应系统作用60 min时,发芽体灭活可达到3.016个对数级。诱导发芽和反应的联合处理程序可显著提高芽胞的灭活水平。     

Production of xylanase from a newly isolated <Emphasis Type="Italic">Penicillium</Emphasis> sp. ZH-30

A new strain of Penicillium sp. ZH-30 that produces xylanase was isolated from soil. According to the morphology and comparison of internal transcribed spacer (ITS) rDNA gene sequence, the strain Penicillium sp. ZH-30 was identified as a strain of Penicillium oxalicum. When xylan or wheat bran was used as substrate at 30°C for 3 days under submerged cultivation, xylanase production was 5.3 and 13.3 U ml⁻¹, respectively. The temperature and pH for optimum activity were 50°C and 5.0–6.0, respectively.     

定量比格犬血浆中去甲文拉法辛LC-MS/MS方法的建立及比格犬血浆中富马酸去甲文拉法辛药代动力学研究

目的:建立一种快速灵敏的液质联用方法定量比格犬血浆中去甲文拉法辛,并进行比格犬体内富马酸去甲文拉法辛药代动力学研究,同时与琥珀酸去甲文拉法辛比格犬体内药代参数进行对比。方法:样品处理采用叔丁基甲醚进行液液萃取,液相分离采用Agilent Eclipse Plus C18色谱柱(2.1 mm×100 mm,3.5μm),以乙腈-乙酸铵缓冲液(1 mmol/L)(体积比80∶20)进行等度洗脱。采用单次给药和多次给药试验进行比格犬体内药代动力学研究。结果:此方法具有良好的线性、精密度、准确度,定量下限(0.5 ng/mL)高于文献报道。液液萃取的样品提取方法简便,液质联用分析时间相对较短(3 min)。结论:此方法可成功地应用于比格犬血浆中去甲文拉法辛定量分析。比格犬体内富马酸去甲文拉法辛的药代参数(单次给药和多次给药试验)与对照品琥珀酸去甲文拉法辛没有明显的统计学差异。     

shRNA靶向干扰COX-2抑制人肝癌裸鼠皮下移植瘤及其血管生成

目的:探讨重组干扰质粒pshRNA-COX-2对人肝癌细胞Hep3B裸鼠皮下移植瘤生长和肿瘤血管生成的抑制作用。方法:重组干扰质粒pshRNA-COX-2转染Hep3B细胞并筛选后,RT-PCR和Western blot检测COX-2mRNA和蛋白表达,RT-PCR检测VEGFmRNA表达。将被成功转染的Hep3B细胞种植于裸鼠皮下,测量肿瘤大小,4周后处死裸鼠,免疫组织化学法检测肿瘤组织中COX-2蛋白表达和肿瘤微血管密度(MVD)。结果:与未转染细胞相比,干扰组COX-2mRNA和蛋白表达抑制率分别为65.3%和52.8%(P<0.05),干扰组VEGFmRNA表达抑制率为56.5%(P<0.05)。干扰组瘤体大小明显小于阴性组和空白组(P<0.01)。干扰组COX-2得分和MVD均明显低于阴性组和空白组(P<0.01)。结论:pshRNA-COX-2通过抑制COX-2表达明显抑制人肝癌细胞Hep3B裸鼠皮下移植瘤生长和肿瘤血管生成。