海南黄牛HSF1 cDNA全长的克隆与序列分析

目的：根据人、小鼠HSF1cDNA保守区序列设计引物,通过PCR方法扩增海南黄牛HSF1cDNA,并进行序列分析。方法：利用RT-PCR、半巢式PCR以及3＇-RACE技术分段扩增得到了海南黄牛HSF1cDNA序列,测序正确后进行拼接。用DNAMAN 生物信息学软件分析海南黄牛HSF1 cDNA与赫里福德牛、人、小鼠同源性和海南黄牛HSF1蛋白的氨基酸组成、等电点、亲/疏水区等蛋白质性质,并根据各种动物HSF1蛋白绘制进化树。结果：（1）海南黄牛的HSF1 cDNA序列全长为1 993bp,包括150bp的5＇非翻译区,1 578bp的开放阅读框以及264bp（不含poly（A）尾）的3＇非翻译区,编码524个氨基酸,分子量为56.42 kD,等电点（pI）为 4.79。（2）海南黄牛的HSF1 cDNA与赫里福德牛、小鼠和人HSF1 cDNA的同源性分别为98.99 %、81.78 %、87.82 %,相应编码蛋白氨基酸序列的同源性分别为98.86 %、83.84 %、89.06 %,其中N-末端和C-末端高度保守,而中间区域存在缺失或替换。（3）根据氨基酸序列构建不同动物HSF1蛋白的进化树,与采用经典遗传分类法构建的进化树基本一致。结论：首次克隆了海南黄牛 HSF1 cDNA全长,分析表明：海南黄牛HSF1蛋白是亲水性蛋白,在8种动物中,其同源性大于73 %,高度保守。海南黄牛与赫里福德牛HSF1蛋白同源性高达98.86 %,在三聚体化区域、转录调节域和激活域存在6个位点的单氨基酸突变,这些发现为进一步揭示海南黄牛抗热性状形成的分子机制提供了重要依据。     

A comprehensive analysis of root morphological changes and nitrogen allocation in maize in response to low nitrogen stress

The plasticity of root architecture is crucial for plants to acclimate to unfavourable environments including low nitrogen (LN) stress. How maize roots coordinate the growth of axile roots and lateral roots (LRs), as well as longitudinal and radial cell behaviours in response to LN stress, remains unclear. Maize plants were cultivated hydroponically under control (4 mm nitrate) and LN (40 μm ) conditions. Temporal and spatial samples were taken to analyse changes in the morphology, anatomical structure and carbon/nitrogen (C/N) ratio in the axile root and LRs. LN stress increased axile root elongation, reduced the number of crown roots and decreased LR density and length. LN stress extended cell elongation zones and increased the mature cell length in the roots. LN stress reduced the cell diameter and total area of vessels and increased the amount of aerenchyma, but the number of cell layers in the crown root cortex was unchanged. The C/N ratio was higher in the axile roots than in the LRs. Maize roots acclimate to LN stress by optimizing the anatomical structure and N allocation. As a result, axile root elongation is favoured to efficiently find available N in the soil.     

Role of the Dinaric Karst (western Balkans) in shaping the phylogeographic structure of the threatened crayfish Austropotamobius torrentium

1. This study examines phylogeography and phylogeny of the threatened stone crayfish, Austropotamobius torrentium, in order to elucidate the role of the Dinaric Karst geology in shaping the evolutionary history and genetic diversity of aquatic fauna in the western Balkans. Mitochondrial 16S rRNA and COI genes were partially sequenced from 188 and 159 crayfish, respectively, sampled from 70 localities. Phylogenetic relationships were reconstructed using four methods of phylogenetic inference. Divergence times between phylogroups were estimated in a Bayesian framework, and their demographic history was examined using neutrality tests and mismatch distribution analysis. 2. Seven geographically localised phylogroups separated by pronounced genetic gaps were found. Five of them have a distribution range within the northern‐central Dinaric (NCD) region, while the remaining two include populations from the southern Balkans (SB) and central and south‐eastern Europe (CSE). The oldest divergence event separated two NCD lineages from the rest of populations in the Late Miocene or Early Pliocene. Divergences amongst the five NCD phylogroups and SB + CSE occurred in the Pliocene. The most recent split separated SB and CSE phylogroups during the Late Pliocene. For both genes, uncorrected pairwise divergences between most of the phylogroups (4.1–8.7% for COI and 1.6–4.8% for 16S rRNA) were of the same range as, or higher than, some of the interspecific distances previously reported for the genus Austropotamobius. 3. Geographically isolated and deeply divergent cryptic monophyletic phylogroups within A. torrentium in the NCD region arose in the course of intensification of Neotectonic movements during the Pliocene and the beginning of the Pleistocene and the development of karstification that has heavily fragmented the palaeohydrography of the area. The results confirm a gradual north–south expansion of stone crayfish during the pre‐Pleistocene that preceded the rapid northward post‐glacial re/colonisation of central Europe (CSE phylogroup) through the Danube drainage. 4. Austropotamobius torrentium comprises morphologically cryptic but molecularly distinct taxa. Considering the relatively small geographical areas they inhabit, the NCD phylogroups of stone crayfish should be given the highest conservation priority.     

1株多重耐药肺炎克雷伯菌耐药基因和整合子、转座子遗传标记研究

目的 了解多重耐药肺炎克雷伯菌的耐药基因存在状况和遗传学背景。方法 聚合酶链反应(RCR)法对多重耐药的肺炎克雷伯菌进行β-内酰胺酶基因、氨基糖苷类修饰酶基因、质粒AmpC酶基因、qacEΔ1-sull耐消毒剂和磺胺基因、整合子遗传标记(整合酶基因)、Tn21/Tn501转座子遗传标记(汞离子还原酶基因)检测。结果 TEM、SHV型β-内酰胺酶基因, DHA型质粒AmpC酶基因,aac(6′)-1型氮基糖苷类修饰酶基因,qacEΔ1-sul1耐消毒剂和磺胺基因,整合子遗传标记(intI1整合酶基因),Tn21/Tn501转座子遗传标记(merA汞离子还原酶基因)检测阳性。结论 多重耐药肺炎克雷伯菌存在多种耐药基因和Ⅰ类整合子、Tn21/Tn501转座子。     

黑土微生物呼吸及群落功能多样性对温度的响应

采用室内培养法,研究了梨树(43°20′N,124°28′E)、德惠(44°12′N,125°33′E)、海伦(47°26′N,126°38′E)和北安(48°17′N,127°15′E)黑土在4℃、15℃和28℃培养条件下的土壤微生物呼吸及Biolog代谢功能多样性.呼吸试验结果表明:尽管土壤微生物呼吸速率在不同培养温度下均表现为北安海伦德惠梨树,但呼吸温度敏感性(Q10值)却随培养温度不同而产生分异,在4℃到15℃变化区间,梨树、德惠、海伦和北安土壤的Q10平均值分别为2.72、3.26、3.21和3.74,而在15℃到28℃变化区间,Q10平均值分别为3.29、2.36、2.11和1.79.不同样点土壤微生物呼吸熵(qCO2)也随培养温度不同而不同,qCO2值在28℃条件下为梨树德惠北安海伦,在15℃条件下为北安德惠海伦梨树,而在4℃条件下样点间变化不大.Biolog试验结果表明:在4℃条件下,微生物群落底物利用碳源数和代谢Shannon指数以海伦和北安黑土较高,而在15℃和28℃条件下,则以梨树和德惠黑土较高.主成分分析表明,海伦与北安、德惠与梨树之间的微生物群落代谢功能相似性较高.综上,不同纬度黑土微生物呼吸特征及群落功能多样性存在差异,高纬度黑土对低温变化敏感,而低纬度黑土对高温变化敏感.     

湖南鱼类新记录4种

2004年8月～2005年7月,在湖南安化进行鱼类资源调查期间,发现湖南鱼类新记录4种：漓江副沙鳅、盎堂拟鲿、越南鲇和司氏（鱼央）.     

药用真菌子实体多糖SEC-HPLC图谱的相似度定量评价研究

目的:对灵芝属Ganoderma下4个种共7个菌株及灰树花Grifola frondosa和云芝Trametes versicolor共9种子实体粗多糖进行了体积排阻HPLC色谱(SEC-HPLC),并分析了相似度.方法:各样品水提物的HPLC图谱用平方欧氏距离法计算了相似度,用离差平方和法进行聚类分析.结果:灵芝属内各样品聚类距离为8,灰树花与云芝的聚类距离为16,它们与灵芝属的距离为25,说明同一灵芝属内的不同种及同种不同菌株所产生的多糖分子量分布近似度远高于来自树花菌属的灰树花或栓菌属的云芝.结论:此方法可量化描述不同多糖间SEC-HPLC的相似关系,有作为真菌多糖样品质量控制手段的潜在能力.     

恶性肿瘤化疗所致院内感染31例分析

目的探讨恶性肿瘤患者化疗后发生院内感染的相关因素及防治措施。方法对31例恶性肿瘤患者化疗后发生的院内感染作回顾性分析。结果恶性肿瘤患者化疗后发生院内感染与中性粒细胞减少程度及持续时间有关,感染的部位以呼吸道为主,主要采用粒细胞集落刺激因子（G-CSF）及抗生素治疗。结论恶性肿瘤患者化疗所致院内感染与中性粒细胞减少程度及持续时间密切相关,合理使用G—CSF及抗生素是预防和控制感染的重要措施。     

肝包虫病的诊断现状及进展

肝包虫病是畜牧地区常见的寄生虫病,是一种人畜共患病,非牧地区偶尔也可以见到,可以通过与狗等动物密切接触直接感染,还可以通过呼吸道吸入虫卵或经消化道误食含有虫卵感染的食物或水源等方式感染使人患病,并且可以寄生于人体内各个部位,但以肝脏最为多见,严重危害人体健康和畜牧业的发展。加强肝包虫病的诊断是防控本疾病的重要措施之一,在不能排除肝包虫病的情况下,应结合病史、化验室检查、B超、CT等影像学检查、免疫学检查进行综合分析,给出明确诊断,以防止误诊、漏诊。近年来随着分子生物学以及病原学等技术的发展,为包虫病的诊断提供更多的方法和信息,本文将从肝包虫病的病史及临床表现、影像学检查、免疫学检查、分子生物学检查及病原学检查等方面对其诊断现状及进展做一综述。     

ω-3多不饱和脂肪酸和三羟异黄酮孕期营养干预对子代大鼠乳腺癌PCNA、ER、BRCA1表达的影响

目的:探讨ω-3多不饱和脂肪酸和三羟异黄酮孕期营养干预对子代大鼠乳腺癌增殖细胞核抗原(PCNA)、雌激素受体(ER)、乳腺癌易感基因1(BRCA1)表达的影响.方法:怀孕大鼠随机分为正常对照组、ω-3多不饱和脂肪酸组、三羟异黄酮低剂量组、三羟异黄酮高剂量组、ω-3多不饱和脂肪酸和三羟异黄酮混合组.选用其雌性子代进行甲基亚硝基脲(MNU)诱导乳腺癌.其中母体为正常对照的分别为子代的正常对照组(Con)和模型组(Mod),其余分组按照母体暴露情况进行划分,分别为ω-3多不饱和脂肪酸组(ω-3PUFA)、三羟异黄酮低剂量组(LGEN)、三羟异黄酮高剂量组(HGEN)、ω-3多不饱和脂肪酸和三羟异黄酮混合组(ω-3PUFA+GEN).观察其乳腺癌发生情况,同时采用RT-PCR和免疫组化检测正常乳腺组织和乳腺癌组织中PCNA、ER和BR-CA1的mRNA或蛋白表达水平.结果:ω-3多不饱和脂肪酸和三羟异黄酮孕期营养干预后可以减少雌性子代MNU诱导乳腺癌的发生,与Con组比较,Mod组乳腺癌组织中BRCA1和ER mRNA表达水平上调,PCNA、ER表达量增加.与Mod组比较,ω-3PUFA组、GEN组乳腺癌组织中BRCA1、ERmRNA及PCNA、ER表达量明显降低.结论:孕期暴露于ω-3PUFA和GEN可降低雌性子代MNU诱导乳腺癌的发生,并下调PCNA、ER和BRCA1的表达量.