大鼠睾丸曲细精管组织支持细胞/生殖细胞长期共培养与精子发生分析

睾丸体外生殖模型的发展为体外研究睾丸的精子发生分子机制和睾丸毒理学提供了实验工具。很多报道的模型都无法真正地模拟体内复杂的生化分子及功能性相互作用从而导致研究价值有限。该实验拟建立一个体外长期维持睾丸生殖细胞存在,并能持续产生精子细胞的支持细胞/生殖细胞共培养体系。体系中的支持细胞和生殖细胞均由曲细精管组织块迁移到培养皿上,在不添加任何生长因子的情况下维持体外精子发生至圆形精子细胞超过2个月。RT-PCR分析显示,共培养细胞稳定表达cdh1、scp3、tnp2;免疫荧光染色结果显示,CDH1、PLZF、SCP3以及SOX9阳性细胞存在。这些结果例证了体系中同时存在精原干细胞、精母细胞、精子细胞和支持细胞。简单高效的支持细胞/生殖细胞体外共培养体系可用于雄性生殖的分子机制和毒理学研究。     

鱼类生殖细胞移植的研究进展

生殖细胞移植在生物学、畜牧学及兴起的鱼类生物工程上都有很多用途。目前,采用基于基因组的育种方法已选育出多种带有所需遗传性状的鱼类新品系;但由于缺乏鱼卵与胚胎的低温保存技术,尚未找到长期保存其种质资源的方法。就鱼类生殖细胞移植的最新研究进展进行综述,同时针对鱼类生殖细胞移植及其低温保存技术研究中存在的问题进行讨论,并提出了鱼类生殖细胞移植的应用前景,以期能积极推动鱼类生物工程的研究,加速鱼类生殖细胞移植在产业中的应用。     

疣粒野生稻应答黄单胞杆菌水稻致病变种（Xoo）的基因芯片

探讨疣粒野生稻应答黄单胞杆菌水稻致病变种（Xoo）的基因芯片制作,通过芯片杂交筛选抗病相关基因。芯片含有2436个片段,来自于应答撖）O的疣粒野生稻差减文库和cDNA文库,通过芯片杂交及微阵列分析基因表达,选其中800个样品点测序比对。其中,35个无同源序列,大部分有同源序列的功能未知,已知功能的序列中明显上调表达的基因有：富含脯氨酸蛋白、泛素连接酶、伸展蛋白、谷胱甘肽S-转移酶II、脂类转移酶等,明显下调表达的基因有：细胞色素P450单加氧酶、醛缩酶、金属硫蛋白、硫氧还蛋白、热激蛋白等,表达无明显变化的基因有：抗坏血酸过氧化物酶、转铜伴侣、脂酶、花丝温敏H2A蛋白等。高通量基因芯片的利用及微阵列分析是筛选抗病相关基因、获取大量抗病相关信息的有效手段。     

葛根素对帕金森病细胞模型的保护作用研究

目的:研究葛根素对帕金森病细胞模型的保护作用及其具体的作用机理.方法:用0.16mM的MPP+处理PC12细胞48h建立帕金森病细胞模型.实验分为对照组、损伤组和保护组,损伤组用MPP+(0.16mM)处理PC12细胞;保护组用葛根素提前预处理PC12细胞1h,后加MPP+.检测PC12细胞存活率、Caspase-3活性及ERβ的转录活性.结果:葛根素能够抑制caspase-3的激活,且其依赖于ERβ的表达,雌激素受体拮抗剂ICI182,780可阻断上述效应;其次葛根素可提高ERβ的转录活性.结论:葛根素对MPP+诱导损伤的PC12细胞具有抗细胞凋亡的保护作用,且具体的作用机理可能依赖于ERβ介导的经典的基因组作用模式.     

锰作用下PC12细胞的氧化应激机制研究

目的：以鼠嗜铬神经瘤细胞（PC12）为模型,筛选锰对神经细胞增殖抑制作用的时间及剂量,观察锰作用下PC12细胞的氧化应激反应与细胞形态学、生化指标改变和丝裂原活化蛋白激酶pp38（p38MAPKs）的活化表达。方法：用200,400,600,800μmol/LMnCl2的培养液,分别作用对数生长期PC12细胞1,2,3,4d后,用MTT筛选锰的细胞毒性剂量;测定200-600μmol/L MnCl2作用4d后,PC12细胞还原型谷胱甘肽和丙二醛含量;透射电镜观察细胞形态学变化;琼脂糖凝胶电泳检测MnCl2对PC12细胞基因组DNA的影响。western-blot法检测p-p38。结果：MTT实验显示200~800μmol/LMnCl2作用1,2,3,4d对PC12有显著的抑制作用,呈剂量和时间依赖趋势,600μmol/LMnCl2作用4d对PC12的抑制率可达50%以上。200-600μmol/LMnCl2作用于细胞4d后,随着浓度的升高,还原型GSH逐渐降低,MDA的含量逐渐升高;600μmol/LMnCl2作用4d电镜可见细胞凋亡,同样条件下细胞DNA碎片化。Western-blot实验显示600μmol/LMnCl2作用1,2,3,4dp-p38逐渐升高,3d时较对照组增加6.6倍（n=3,p〈0.05）,200,400,600μmol/L MnCl2作用4d时,磷酸化蛋白38（p-p38）也逐渐升高,400μmol/L MnCl2作用4d时较对照组升高了4.7倍（n=3,p〈0.05）。结论：PC12细胞在锰作用下发生氧化应激反应,上调p-p38,诱导细胞凋亡,细胞增殖抑制。     

血浆B型脑钠肽对心力衰竭和呼吸困难的诊断价值

目的：探讨血浆脑钠肽（BNP）水平在心力衰竭（CHF）T和呼吸困难诊断中的应用。方法：采用免疫荧光快速测试法测定56例已确诊心衰患者、40例心源性呼吸困难患者、29例肺源性呼吸困难患者和30例健康人血浆BNP的含量。结果：心衰组患者血浆BNP水平明显高于对照组,差异显著（P〈0．01）;心源性呼吸困难组患者的BNP值水平（1032．2±879．8pg／ml）与肺源性呼吸困难组患者的BNP值水平（67．1±43．6pg／ml）比较有显著性差异（P〈0．01）。结论：检测BNP水平可为临床诊断CHF及心源性与肺源性呼吸困难的鉴别诊断提供重要依据。     

一头遭鲨鱼袭击和误捕的小须鲸幼仔

A minke whale was by-caught by fishermen in a drift gill net off Shidao, Weihai, Shandong Province on June 15, 2007. The whale, which had been attacked by a shark, was a male with body length 2.43 m, and was suspected to have died soon after the birth. The small size of the whale suggests that a breeding ground for minke whales might exist in the Chinese coastal waters. Evidence of shark attack on this young whale was apparent: half of the fluke was lost and the belly was bitten as well; injuries were not as severe on the caudal peduncle. It was not possible to determine if the shark-related injuries observed occurred prior to the whale being by-caught or were post-mortem. Some conservation measures were proposed in order to better protect the cetaceans in waters off China.     

硅酸盐细菌HM8841菌株解钾作用的研究

早在三十年代苏联学者从土壤中分离到硅酸盐细菌,并测定解钾强度,培养5d分离出来的钾量为原硅酸盐中含量的15.9％,同时在不同土壤和不同农作物上进行了盆栽和田间试验,证明对农作物生长有较好的促进作用和增产效果。也有一些学者认为硅酸盐细菌的作用是刺激作用,为农作物补充钾素营养是微不足道的。 作者分离到硅酸盐细菌HM8841菌株,经工业发酵研制成菌剂。该菌剂经几年的大面积推广应用,在缺钾土壤上对各种农作物均表现出较好的增产效果。尤其在土壤中速效钾严重不足的情况下,施用该菌剂增产更加明显。这充分说明土壤缺少速效钾是限制农作物增产的重要原因。HM8841菌株能为农作物提供一定量的钾素使农作物提高产量。但其解钾能力有多高,能提供多少钾素,需要进一步研究。为此作者采取不同的方法,不同的底物对HM8841菌株的解钾效能进行了研究。     

Gelatin degradation assay reveals MMP-9 inhibitors and function of O-glycosylated domain

AIM: To establish a novel, sensitive and high-throughput gelatinolytic assay to define new inhibitors and compare domain deletion mutants of gelatinase B/matrix metalloproteinase (MMP)-9. METHODS: Fluorogenic Dye-quenched (DQ)TM-gelatin was used as a substrate and biochemical parameters (substrate and enzyme concentrations, DMSO solvent concentrations) were optimized to establish a highthroughput assay system. Various small-sized libraries (ChemDiv, InterBioScreen and ChemBridge) of hetero-cyclic, drug-like substances were tested and compared with prototypic inhibitors. RESULTS: First, we designed a test system with gelatin as a natural substrate. Second, the assay was validated by selecting a novel pyrimidine-2,4,6-trione (barbitu- rate) inhibitor. Third, and in line with present structural data on collagenolysis, it was found that deletion of the O-glycosylated region significantly decreased gelatinolytic activity (kcat/kM ± 40% less than full-length MMP-9). CONCLUSION: The DQTM-gelatin assay is useful in high-throughput drug screening and exosite targeting. We demonstrate that flexibility between the catalytic and hemopexin domain is functionally critical for gelatinolysis.     

Ellenberg indicator values for macromycetes – a methodological approach and first applications

Ellenberg indicator values (EIVs) describe the realized niche of species and habitat parameters, and are commonly used for vascular plants, bryophytes, and lichens. We provide a methodology for EIVs and an EIV list for nearly 650 species of macromycetes. We propose a new EIV scale, namely substrate openness (O). We also give the results of two applications and compare EIV values related to the Red List classification with those related to lifestyle classification. Mycorrhizal species on average have higher demands on substrate openness and are less tolerant of high nutrient levels than saprotrophic or parasitic species. Critically endangered species have on average distinctly higher demands on openness of habitat and substrate than not threatened or less strongly threatened species, which in turn have higher demands for nutrient availability. This pattern clearly highlights the points of threat for many macromycete species.