斜卧青霉原生质体制备和再生的研究

研究了斜卧青霉原生质体制备和再生的条件,确定了培养方式、菌龄、渗透压稳定剂、酶的配比、酶解时间等因素对原生质体制备和再生的影响。最佳条件：菌丝体培养12h,用1％溶壁酶＋1％纤维素酶＋1％蜗牛酶的混合酶液酶解,将NaCl作为渗透压稳定剂,酶解时间为1．5h。在此条件下原生质体的形成量达到5．3×10^5个／mL,再生率为19．4％。     

李拭库蠓生活史的研究(双翅目:蠓科)

在重庆地区所作有关李拭库蠓生活史的研究,观察在实验条件结合自然情况下进行。设计了虫尸培养法,在虫尸培养内幼期处于严格控制的条件下,可以确切地观察卵的孵化、幼虫蜕皮、化蛹和羽化等现象,解决了库蠓变态发育的规律。 李拭库蠓的主要动物宿主是牛和家猪,雌虫吸取血液后糖类对卵巢的发育并不是必需的。9—11.5℃时胃血消化和卵巢发育仍能缓慢地进行。雌虫在20—28℃时一般在吸血后第3—4日间产卵,产卵数为40—213。 幼期发育的时间随温度和食物而改变。虫尸培养,在27±1℃时自卵至成虫为28—44天(卵期3天:第一龄幼虫6—8天:第二龄幼虫5—10天;第三龄幼虫6—9天:第四龄幼虫5一11天;蛹期3天)。当6.8—22℃时,共需101—212天(卵期3—8天;第一龄幼虫9—31天;第二龄幼虫27—35天;第三龄幼虫34—48天:第四龄幼虫25—80天:蛹期3—10天)。夏季室温条件下泥土培养基内自卵至成虫为22—32天。     

Loss of decreased-rubisco phenotype between generations of wheat transformed with antisense and sense rbcS

The elite UK winter wheat cv. Riband was transformed with constructs containing rbcS in sense and antisense orientations driven by the maize ubiquitin promoter with a transformation efficiency of 1.2%. Of 77 primary transformants 31% of the sense-rbcS transformed lines and 78% of the antisense-rbcS transformed lines had decreased rubisco content compared to wild-type and marker-only controls, with decreases of up to 60%. However, in the T1 progeny which inherited the transgene, only 5% showed significantly decreased rubisco content and these effects were on the margins of significance. Five potential T2 homozygous lines from T1 parents which had transgene segregation consistent with a single locus were identified. There was no significant decrease in rubisco content relative to wild-type in any of these lines (LSD of 8% for P= 0.05). Expression of antisense rbcS transgenes in two of these T2 lines was low but was increased following exposure of the plants to 37°C for 48 h. However this did not induce a significant decrease in rubisco protein content relative to controls. Southern analysis of two antisense lines showed that they had low copy number and 1–2 insertion events. In one of the two lines there was increased methylation of the ubiquitin intron in T2 samples compared to the TO primary transformant. Further work is required to establish whether methylation occurred in all the lines which lost the phenotype, and therefore the likelihood of this being the cause. The disappearance of the decreased rubisco-content phenotype between generations may therefore be attributable to (1) greater activity of the ubiquitin promoter due to greater stress in the T0 generation plants and/or (2) increased methylation of the transgene promoter region between generations.     

Genome-Scale Screening of Drug-Target Associations Relevant to Ki Using a Chemogenomics Approach

The identification of interactions between drugs and target proteins plays a key role in genomic drug discovery. In the present study, the quantitative binding affinities of drug-target pairs are differentiated as a measurement to define whether a drug interacts with a protein or not, and then a chemogenomics framework using an unbiased set of general integrated features and random forest (RF) is employed to construct a predictive model which can accurately classify drug-target pairs. The predictability of the model is further investigated and validated by several independent validation sets. The built model is used to predict drug-target associations, some of which were confirmed by comparing experimental data from public biological resources. A drug-target interaction network with high confidence drug-target pairs was also reconstructed. This network provides further insight for the action of drugs and targets. Finally, a web-based server called PreDPI-K_i was developed to predict drug-target interactions for drug discovery. In addition to providing a high-confidence list of drug-target associations for subsequent experimental investigation guidance, these results also contribute to the understanding of drug-target interactions. We can also see that quantitative information of drug-target associations could greatly promote the development of more accurate models. The PreDPI-K_i server is freely available via: http://sdd.whu.edu.cn/dpiki.     

大鼠细小病毒H-1株培养方法的建立

目的建立用大鼠胶质瘤细胞系（Rat glial cell line C6）替代大鼠原代胚细胞（primary rat embryocells,RE）培养大鼠细小病毒（Toolan virus,H-1）的方法。方法 将0.8 mL H-1病毒接种C6细胞（75T培养瓶,细胞接种量为2×105/mL,培养过夜）,待细胞病变CPE达＋＋～＋＋＋时,用免疫荧光（FITC）鉴定所培养病毒的抗原,用血球凝集试验（HA）测定培养物上清效价,用DNA测序鉴定所培养的病毒,用96孔板培养法测定H-1病毒的TCID50。结果H-1在接种到C6细胞的第3～4天,细胞发生明显的病变,CPE可达＋＋＋＋,FITC鉴定呈H-1抗原阳性,病毒的培养上清中HA效价为1∶320。测序结果表明：该病毒序列与NCBI中H-1序列同源性达99%,确定为H-1病毒。收获的H-1的TCID50为103.2/0.1 mL。结论用大鼠胶质瘤细胞系（C6）可以替代大鼠原代胚细胞（RE）进行大鼠细小病毒的培养。     

贵州师范大学植物标本室的历史与沿革

自2017年参加“国家标本资源共享平台”植物子平台以来,贵州师范大学地理与环境科学学院植物标本室除了数字化植物标本以外,还挖掘、整理了贵州师范大学植物分类学、植物地理学的建设和发展历程。经过3年的艰苦努力,最后向中国数字植物标本馆(CVH)提供了10 044份信息齐全的植物标本。通过此次植物标本数字化工作,彻底摸清了贵州师范大学地理与环境科学学院植物标本的家底,盘活了标本室的库存,使得老一辈植物地理学家黄威廉等老先生60余年的科研成果重新展现在世人面前。通过植物标本的数字化工作,挖掘整理出老一辈精彩的植物标本采集史和学术故事。在标本数字化的过程中,学生和老师都对植物标本以及植物标本背后的故事有了新的认识,对植物标本数字化的重要性和必要性也有了更深的体会。     

比较两种测量方法获得的血压波动对老年颈动脉斑块的影响

目的:比较两种不同血压波动测定方法测定的血压波动与颈动脉斑块发生的关系。方法:以1456名患有动脉硬化老年男性患者为研究对象,监测患者24 h动态血压,根据有无颈动脉斑块将入选患者分为2组:颈动脉斑块组(n=1012)和无颈动脉斑块组(n=444),分别采用经典的标准差方法(SD法)和个体血压波动测定方法(个体法)分别测定每位患者的血压波动,回顾性分析这两种方法测定的血压波动与颈动脉斑块形成的关联性。结果:SD法测定颈动脉斑块组24 h收缩期血压波动(SBPV)、白天SBPV、夜间SBPV以及24 h舒张期血压波动(DBPV)水平均明显高于无颈动脉斑块组(P0.05);而白天和夜间DBPV差异无统计学意义。个体法测定颈动脉斑块组24 h SBPV、白天SBPV、24 h DBPV以及白天DBPV水平较无颈动脉斑块组均明显升高(P0.05);夜间SBPV和夜间DBPV差异无统计学意义(P0.05)。比较颈动脉斑块组SBPV值出现次数,SD法测定SBPV最多的是10-15 mmHg(n=541),其次是大于15 mmHg(n=399);个体法测定颈动脉斑块组SBPV值出现次数最多的是0-8 mmHg(n=490),其次是8-10 mmHg(n=350)。结论:在老年男性动脉硬化相关疾病患者中,血压波动与颈动脉斑块的形成有着密切的关系,两种方法均可测定血压波动,但以个体血压波动测定方法更加敏感。     

水稻PCR扩增模板的快速制备

用碱处理水稻嫩叶,取碱处理水稻叶片浸出液直接用作PCR扩增的模板,扩增结果稳定、准确,与常规提取的DNA扩增效果相比没有显著差异。用此方法制备的模板室温下2周之内、4℃下3周之内、-20℃下4个月以上扩增结果不变。此方法所需试剂均为常规试剂,具有需要材料少、成本低、简便、快捷等优点,尤其适合材料比较宝贵的转基因水稻的预筛选和大样本量的PCR检测,为分子标记技术的实际应用创造了条件。 Abstract:The solution of alkali-treated fresh rice leaves was used directly as the templates of PCR.The amplified results were stable,reliable,and had no difference compared with that amplified with rice total DNA extracted by common method.The stable results can still be obtained based on the templates kept at 25℃ for tow weeks,at 4℃ for three weeks,at -20℃ for over four months.With this technique,less material and only common reagent are required,which is especially adapted to the screening of the precious trangenic rice in advance and large-scale PCR tests.     

苦草属植物雄花果实种子显微结构的研究

苦草属植物体外部形态相似,叶线行或带形,野外难以直接鉴定到种,而雄花雄蕊数目、果实和种子的显微结构是苦草属(Vallisneria)植物物种的鉴别性特征之一。苦草属物种的雄花小,仅有1.5 mm,人工解剖雄花观察方法难以实现雄花雄蕊数目的识别;本研究采用雄株雄佛焰苞水培预培养技术获取自然开放的雄花并利用解剖学显微镜对3种苦草属植物雄花、果实和种子进行了解剖观察和研究。显微结构观察表明,苦草(V. natans)雄蕊1枚,果实圆柱形,表面光滑,种子无翅,具纵条纹;刺苦草(V. spinulosa)雄蕊2枚,果实三棱形,棱上具刺,种子具翅;密刺苦草(V. denseserrulata)雄蕊2枚,果实三棱圆柱形,表面光滑,种子无翅。依据文献,密刺苦草、刺苦草、长梗苦草(V. longipedunculata)和安徽苦草(V. anhuiensis)是安徽苦草属新分布记录种,其中长梗苦草和安徽苦草是分布于安徽的新种。     

宏转录组测序分析老年抗生素相关性腹泻患者肠道菌群组成及耐药通路特点

为探究老年抗生素相关性腹泻(AAD)患者肠道菌群结构变化的特点以及与细菌耐药性之间的关联。

对来自华山医院老年科的6名老年AAD患者的肠道菌群进行宏转录组测序, 并以另5名相同饮食标准下接受抗生素治疗但未发生腹泻的患者作为对照, 深入分析老年AAD患者肠道菌群物种组成及耐药通路变化。

AAD组物种丰度和多样性均低于对照组, 且两组菌群组成在不同分类学水平上存在明显差异(P < 0.05)。KEGG比对结果显示, AAD组多条抗生素耐药通路的表达量均有显著上调, 包括与β-内酰胺类抗生素耐药相关的青霉素结合蛋白pbp1B、pbp5、金属β-内酰胺酶bla2, β-内酰胺酶基因表达调节因子ampR、ampC等, 与万古霉素耐药相关的VanX、VanB、VanW和VanH等。另外, mlaC、natB、znuC、tcyB、tcyC等53条ATP结合盒超家族(ABC转运蛋白)相关通路在AAD组中显著上调。

菌群结构变化、某些优势菌属的扩张、菌群失调相关的代谢变化以及抗生素耐药性等因素与老年人AAD有潜在联系。