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。     

LATS1过表达对肺癌A549细胞增殖及周期的影响

通过过表达手段上调大肿瘤抑制因子1（1arge tumor suppressor gene 1,LATS1）基因在A549细胞中的表达,研究LATS1对A549细胞生长和细胞周期调控的作用。构建过表达LATS1基因的慢病毒载体,转染A549N胞株,采用RT-PCR和蛋白质印迹法检测转染后A549细胞中LATS1、YAPmRNA和蛋白的表达效率;流式细胞术检测细胞凋亡、周期情况：CCK-8检测细胞的增殖水平变化。结果发现,过表达LATS1慢病毒载体转染A549细胞株后,LATS1mRNA及蛋白表达水平高于未处理组及转染空载体组,YAPmRNA及蛋白表达水平低于未处理组及转染空载体组;过表达LATS1慢病毒转染后,A549细胞增殖率从第五天开始低于对照组（P〈0．05）,过表达组细胞G1期比例明显增高（P〈0．05）,凋亡率明显增加（P〈0．05）,差异均有统计学意义。以上结果提示,LATS1可通过下调YAP的表达水平促进A549细胞的凋亡,诱导G1期阻滞,降低细胞的增殖能力。     

肝炎平对D－氨基半乳精所致肝损害保护作用的酶组织化学研究

本实验采用Ｄ－氨基半乳糖（Ｄ－ＧａｌＮ）诱导的大鼠急性肝损伤模型,观察大鼠肝脏组织化学的变化,探讨肝炎平对急性肝损伤的保护作用。实验分为四组,即正常对照组、模型组、肝炎平及肝得健保护组。结果表明：肝炎平对肝细胞膜系统有一定的保护作用。肝炎平组和肝得健组ＳＤＨ、ＣＣＯ及ＣｈＥ活性明显高于模型组,且与正常对照组相近。本实验模型组ＡＣＰ的活性明显高于正常组,而肝炎平组ＡＣＰ的活性明显低于模型组,与正常对照组无显著性差异。提示：肝炎平可显著改善因Ｄ－氨基半乳糖所致肝损害的作用。且其对肝细胞的保护作用与肝得健一致。     

椎体静脉稀疏区注入骨水泥对PVP术中骨水泥渗漏的影响

目的:探讨椎体静脉稀疏区注入骨水泥对骨质疏松椎体压缩性骨折患者行经皮穿刺椎体成形术(percutaneous vertebroplasty,PVP)术中骨水泥渗漏的影响。方法:选择西安交通大学第二附属医院2014年1月至2018年6月收治的61例骨质疏松椎体压缩性骨折患者,根据骨水泥注入区域的不同,将所有患者分为A组(30例)及B组(31例),A组骨水泥注入区域为椎体静脉密集区(椎体中1/3平面处),B组骨水泥注入区域为椎体静脉稀疏区(椎体上1/3及下1/3平面处),对比两组的骨水泥渗漏率,术前、术后6个月时的视觉模拟评分(Visual analogue scale,VAS),治疗中的骨水泥用量、椎体高度恢复率及cobb角恢复度数。结果:B组的骨水泥渗漏率及骨水泥用量均明显低于A组(P0.05)。两组的VAS评分、椎体高度恢复率、cobb角恢复情况对比差异无统计学意义(P0.05)。结论:与椎体静脉密集区相比,在椎体静脉稀疏区注入骨水泥可显著降低骨质疏松椎体压缩性骨折患者PVP术中骨水泥渗漏率,椎体静脉稀疏区可作为PVP术中骨水泥注射的一个相对安全区域。     

Physical mapping of the 5S and 45S rDNA in teosintes

The physical locations of the 5S and 45S rDNA sequences were examined in three types of teosinte, Zea mays ssp. mexicana (2n = 20), Zea diploperennis (2n = 20) and Zea perennis (2n = 40) by biotinylated fluorescence in situ hybridization (FISH). The tested materials only showed one hybridization site of 5S rDNA on their genomes, but they were different in the position of the signals. The hybridization site of Zea mays ssp. mexicana was located on the long arm of chromosome 2, indicating that it is the same as the cultivated maize in the position of 5S rDNA, while the sites of Zea diploperennis and Zea perennis were on the short arms of other chromosomes. For 45S rDNA, one hybridization site was detected at secondary constriction region of the satellite chromosomes in Zea mays ssp. mexicana and Zea diploperennis, while in Zea perennis, besides the site located at the secondary constriction region, a second site on the short arm of another chromosome pair was observed. Our results provide additional evidence for Zea mays ssp. mexicana being a subspecies of Zea mays.     

川中丘陵地区大豆根瘤菌遗传多样性与系统发育关系

【目的】研究分离自川中丘陵地区大豆根瘤菌的遗传多样性和系统发育。【方法】采用16S rDNA PCR-RFLP和16S rRNA基因、glnII、共生基因(nodC)系统发育分析的方法进行研究。【结果】供试未知菌的16S rDNA用4种限制性内切酶(HaeⅢ、HinfⅠ、MspⅠ及TaqⅠ)酶切后获得5种16S遗传图谱类型。16S rDNA PCR-RFLP结果表明,所有供试菌株在83%水平分为慢生根瘤菌属(Bradyrhizobium)和中华根瘤菌属(Sinonrhizobium)两大类群,而75%的菌株为中华根瘤菌。6个代表菌株的16S rDNA、glnII和nodC三个位点基因的系统发育结果基本一致,4株与S.fredii USDA205T相似度最高;有2株分别与B.yuanmingense CCBAU10071T、B.diazoefficiens USDA110T相似度最高。4个Sinonrhizobium代表菌株16S rDNA、glnII序列相似度分别为98.3%-99.9%、98.2%-100%,但它们的nodC基因序列完全相同。【结论】川中丘陵地区大豆根瘤菌具有较丰富的遗传多样性,S.fredii为优势种。     

濒危植物明党参与非濒危种峨参种子休眠和萌发比较

研究了濒危植物明党参（Changium smyrnioides)与非濒危种峨参（Anthriscus sylvestris)种子贮存,打破休眠和萌发对水分和温度条件的要求。结果表明,在自然条件下,明党参种子有5个月的休眠期,人工低温（10℃左右）处理40天即可打破休眠;两种植物种子自然温度干燥处理不能打破休眠;两种植物的种子在自然温度变湿层积处理后萌发率最高,萌发持续时间也最长,其中明党参的萌发率地峨参,持续时间短于峨参;自然温度淹水处理大大降低了两种植物种子的萌发率,但明党参仍有7％的萌发率。明党参种子质量和发芽率不应是明党参濒危的直接原因,但因其具有种子产量低,幼苗数量少,存活率高的K－对策,当受到强烈干扰时,种群难以在短期内恢复,容易濒危。     

Directed differentiation of induced pluripotent stem cells towards T lymphocytes

Adoptive cell transfer (ACT) of antigen-specific CD8(+) cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of malignancies (1). CTLs can recognize malignant cells by interacting tumor antigens with the T cell receptors (TCR), and release cytotoxins as well as cytokines to kill malignant cells. It is known that less-differentiated and central-memory-like (termed highly reactive) CTLs are the optimal population for ACT-based immunotherapy, because these CTLs have a high proliferative potential, are less prone to apoptosis than more differentiated cells and have a higher ability to respond to homeostatic cytokines (2-7). However, due to difficulties in obtaining a high number of such CTLs from patients, there is an urgent need to find a new approach to generate highly reactive Ag-specific CTLs for successful ACT-based therapies. TCR transduction of the self-renewable stem cells for immune reconstitution has a therapeutic potential for the treatment of diseases (8-10). However, the approach to obtain embryonic stem cells (ESCs) from patients is not feasible. Although the use of hematopoietic stem cells (HSCs) for therapeutic purposes has been widely applied in clinic (11-13), HSCs have reduced differentiation and proliferative capacities, and HSCs are difficult to expand in in vitro cell culture (14-16). Recent iPS cell technology and the development of an in vitro system for gene delivery are capable of generating iPS cells from patients without any surgical approach. In addition, like ESCs, iPS cells possess indefinite proliferative capacity in vitro, and have been shown to differentiate into hematopoietic cells. Thus, iPS cells have greater potential to be used in ACT-based immunotherapy compared to ESCs or HSCs. Here, we present methods for the generation of T lymphocytes from iPS cells in vitro, and in vivo programming of antigen-specific CTLs from iPS cells for promoting cancer immune surveillance. Stimulation in vitro with a Notch ligand drives T cell differentiation from iPS cells, and TCR gene transduction results in iPS cells differentiating into antigen-specific T cells in vivo, which prevents tumor growth. Thus, we demonstrate antigen-specific T cell differentiation from iPS cells. Our studies provide a potentially more efficient approach for generating antigen-specific CTLs for ACT-based therapies and facilitate the development of therapeutic strategies for diseases.     

杆状病毒转导不同哺乳动物骨髓来源间充质干细胞

杆状病毒作为一种新型基因载体,若能有效转导不同哺乳动物骨髓来源间充质干细胞(bone marrow-derived mesenchymal stem cells, BMSCs),将会成为干细胞基因修饰研究领域中更理想的一种基因载体.本文探讨了重组杆状病毒(BacV-CMV-EGFP)对不同哺乳动物BMSCs的转导效率.体外原代培养小鼠、大鼠、猪、恒河猴及人的BMSCs.用培养3代以上的哺乳动物BMSCs进行病毒转导实验,转导2d后用倒置荧光显微镜观察绿色荧光蛋白在不同哺乳动物BMSCs中的表达,并用流式细胞仪检测重组杆状病毒对不同哺乳动物BMSCs的转导效率.结果显示:原代培养的小鼠、大鼠、猪、恒河猴及人的BMSCs于体外传代3次以上后,细胞呈现较均一的梭形,漩涡状生长;倒置荧光显微镜观察显示,与小鼠、大鼠、猪的BMSCs相比,恒河猴及人有更多BMSCs表达绿色荧光蛋白,且荧光强度较强;杆状病毒对小鼠、大鼠、猪、恒河猴及人的BMSCs的转导效率分别为(21.21±3.02)%、(22.51±4.48)%、(39.13±5.79)%、(71.16±5.36)%及(70.67±3.74)%.上述结果表明,重组杆状病毒对不同哺乳动物BMSCs的转导效率不同,对恒河猴及人的BMSCs转导效率较高,说明重组杆状病毒可作为人或灵长类动物BMSCs基因修饰研究领域中更理想的基因载体.