生态袋护坡技术在三峡水库消落带植被恢复中应用的可行性研究

通过对三峡水库重庆市巫山县双龙镇和巫峡镇段消落带开展生态袋护坡复绿试验7年后，生态袋上（内）、生态袋堆叠处上方和左侧消落带狗牙根（Cynodon dactylon）的种群密度、表型生长性状、地上和地下生物质量，以及土壤理化性质的测定，探讨以狗牙根为生态袋上的种植植物，将生态袋护坡技术用于三峡水库消落带植被恢复的可行性。结果表明：（1）各试验地生态袋上与其堆叠处上方和左侧消落带的狗牙根种群密度和地上生物质量差异不显著。（2）狗牙根的表型生长性状和根系生物质量因地和在生态袋堆叠处的方位不同而异。在双龙镇试验地，生态袋上比其堆叠处上方消落带上狗牙根的植株长度和节间长度低23.9%和22.6%（P<0.05），除此之外的各项指标差异均不显著；生态袋内0-5 cm土层的根系生物质量比其堆叠处上方消落带增加了75.7%（P<0.05），比其堆叠处左侧消落带降低了11.8%，在5-15 cm各土层降低了91.6%-96.9%（P<0.05），15-20 cm土层的差异不显著。在巫峡镇试验地，生态袋上与其堆叠处上方和左侧消落带的各表型生长性状的差异均不显著；生态袋内各土层的根系生物质量均比其堆叠处上方和左侧消落带增加了20.0%-138.7%。（3）各试验地生态袋内与其堆叠处上方和左侧消落带土壤容重的差异不显著，土壤化学性质因地和在生态袋堆叠处的方位不同而异。在双龙镇试验地，生态袋内的土壤全氮和速效氮含量比其堆叠处上方消落带分别降低了13.6%和40.9%（P<0.05），比其堆叠处左侧消落带分别降低了11.9%和33.0%（P<0.05）；速效钾含量比其堆叠处上方和左侧消落带分别增加了18.3%和34.1%（P<0.05）；除此之外各指标的差异均不显著。在巫峡镇试验地，生态袋内的土壤pH值和全氮含量比其堆叠处上方消落带分别降低了1.4%和27.9%（P<0.05），全钾含量增加了6.1%（P<0.05）；土壤全钾和速效钾含量比生态袋堆叠处左侧消落带分别降低了8.1%和24.9%（P<0.05）；除此之外各指标的差异也不显著。（4）狗牙根种群密度、大多数生长指标和生物质量与土壤理化指标相关不紧密。总体上，生态袋上（内）与其堆叠处上方和左侧消落带的大多数表型生长指标，地上和地下生物质量，以及土壤理化指标的差异不显著。狗牙根耐淹、抗旱、耐贫瘠，根系发达，且穿透力强，能够在生态袋上正常生长；生态袋透水不透土，且具有一定的保肥能力。因此，以狗牙根为生态袋上的种植植物，将生态袋护坡技术用于三峡水库消落带植被恢复具有一定的可行性。     

HCMV IE86对恶性胶质瘤细胞凋亡及p53表达活性的影响

人巨细胞病毒(human cytomegalovirus,HCMV)能诱导肿瘤细胞恶性转化且抑制肿瘤细胞凋亡,但HCMV编码的主要即刻早期调控蛋白IE86在这一过程中是否发挥关键作用仍然未知。为探究IE86对基因修饰荷胶质瘤小鼠p53表达水平及恶性胶质瘤细胞凋亡情况的影响,本研究通过PCR技术鉴定基因修饰小鼠IE86表达情况;实时定量PCR技术检测IE86和p53mRNA表达水平变化;免疫组织化学方法检测p53和p21蛋白的表达水平;TUNEL检测肿瘤组织细胞凋亡情况。结果显示,成功构建了IE86基因修饰小鼠模型;与IE86阴性组相比IE86阳性组p53表达水平上升(P0.05),但p21表达水平下降(P0.05);IE86阳性组细胞抗凋亡能力增强(P0.05)。以上结果表明,在基因修饰的小鼠中IE86持续表达但p53转录活性的指示标志p21下调,且IE86可提高恶性胶质瘤细胞抗凋亡能力。     

机械刺激对匍匐茎型植物蛇莓(Duchesnea indica)克隆可塑性的影响（英文）

机械刺激(mechanical stimuli, MS)在自然界的分布较为普遍,然而关于机械刺激对匍匐茎型克隆植物影响的研究相对较少。为此,实验对匍匐茎型克隆植物蛇莓(Duchesneaindica)进行了对照(无机械刺激)、克隆片段顶部刺激、半克隆片段刺激和整克隆片段刺激共四个水平的机械刺激处理,通过研究蛇莓克隆生长的响应,探讨机械刺激对其作用机理。结果表明:与对照相比,机械刺激导致蛇莓产生较多的分株、较多的匍匐茎、短而细的叶柄及较多的根生物量分配。同时,不同作用位点的机械刺激对各生理生长指标的效应差异并不显著。另外,通过对各生长指标表型可塑性指数的分析发现,分株数具有较高的可塑性指数。上述实验结果表明,蛇莓可以较好地适应机械刺激干扰的生境,其分株数可以作为其响应程度的较好的指示指标。     

油茶林鳞翅目害虫3种天敌记述

采用林间调查和室内饲养等方法,发现了油茶林鳞翅目害虫的3种天敌,即印度长颈步甲Ophionea indica、日本长颈步甲Mimocolliuris insulana和茶梢尖蛾长体茧蜂Macrocentrus parametriates ivorus,前2种天敌主要捕食鳞翅目卷叶类害虫、假眼小绿叶蝉Empoasca vitis等,且为首次在油茶林中发现;茶梢尖蛾长体茧蜂寄生茶梢尖蛾Parametriotes theae幼虫,为首次在广东省记录。记述了3种天敌的形态特征、寄主、国内分布及生物学特性等。     

The porcine chloride channel calcium-activated family member pCLCA4a mirrors lung expression of the human hCLCA4

Pig models of cystic fibrosis (CF) have recently been established that are expected to mimic the human disease closer than mouse models do. The human CLCA (originally named chloride channels, calcium-activated) member hCLCA4 is considered a potential modifier of disease severity in CF, but its murine ortholog, mCLCA6, is not expressed in the mouse lung. Here, we have characterized the genomic structure, protein processing, and tissue expression patterns of the porcine ortholog to hCLCA4, pCLCA4a. The genomic structure and cellular protein processing of pCLCA4a were found to closely mirror those of hCLCA4 and mCLCA6. Similar to human lung, pCLCA4a mRNA was strongly expressed in porcine lungs, and the pCLCA4a protein was immunohistochemically detected on the apical membranes of tracheal and bronchial epithelial cells. This stands in sharp contrast to mouse mCLCA6, which has been detected exclusively in intestinal epithelia but not the murine lung. The results may add to the understanding of species-specific differences in the CF phenotype and support the notion that the CF pig model may be more suitable than murine models to study the role of hCLCA4.     

Binding of trastuzumab to ErbB2 is inhibited by a high pericellular density of hyaluronan

Although trastuzumab is an efficient drug, primary and acquired resistance is a challenging problem. The authors have previously shown in mouse xenograft experiments that masking ErbB2 by hyaluronan leads to diminished binding of the antibody and consequent resistance. In the current work, they correlated trastuzumab binding with the pericellular density of hyaluronan in ErbB2-overexpressing human breast cancer samples. A method for quantifying the relative binding of trastuzumab was developed involving constant and low-frequency background subtraction, segmenting the image to membrane and background pixels followed by evaluation of trastuzumab fluorescence, normalized with the expression level of ErbB2, only in the membrane. The normalized binding of trastuzumab showed a negative correlation with the pericellular density of hyaluronan (r = -0.52) with the effect being the most pronounced in the extreme cases (i.e., low and high hyaluronan densities predicted strong and weak binding of trastuzumab, respectively). Removal of hyaluronan by hyaluronidase digestion unmasked the trastuzumab binding epitope of ErbB2 demonstrated by a significantly increased normalized binding of the antibody. The results show that the accumulation of pericellular hyaluronan plays a crucial role in masking ErbB2.     

YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers

The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3β, PDX1, CD34, p63, nestin, PAX6) markers. Double-labeling showed YKL-40 expression in OCT4-positive hESCs, PAX6-positive neuroectodermal cells, and HNF-3β-positive endodermal cells. The differentiating progeny showed strong YKL-40 expression. Abrupt transition between YKL-40 and OCT4-positive hESCs and YKL-40-positive ecto- and neuroectodermal lineages was observed within the same epithelial-like layer. YKL-40-positive cells within deeper layers lacked contact with OCT4-positive cells. YKL-40 may be important in initial cell differentiation from hESCs toward ectoderm and neuroectoderm, with retained epithelial morphology, whereas later differentiation into endoderm and mesoderm involves a transition into the deeper layers of the colony.     

Effect of reactivity on cellular accumulation and cytotoxicity of oxaliplatin analogues

The purpose of this study was to systematically investigate the relationships between reactivity, cellular accumulation, and cytotoxicity of a panel of oxaliplatin analogues with different leaving groups in human carcinoma cells. The reactivity of the complexes towards the nucleotides 2'-deoxyguanosine 5'-monophosphate and 2'-deoxyadenosine 5'-monophosphate was studied using capillary electrophoresis. Cellular accumulation and cytotoxicity were measured in an oxaliplatin-sensitive and oxaliplatin-resistant ileocecal colorectal adenocarcinoma cell line pair (HCT-8/HCT-8ox). Platinum concentrations were determined by flameless atomic absorption spectrometry. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to assess cytotoxicity. Early cellular platinum accumulation was predominantly affected by lipophilicity. A relationship between reactivity and cellular accumulation was observed for three of four platinum complexes investigated, whereas the most lipophilic oxaliplatin analogue was an exception. Increased reactivity and reduced lipophilicity were associated with high cytotoxic activity. Resistance was influenced by lipophilicity but not by reactivity. The observed relationships may help in the design of analogues with high antitumoral activity in oxaliplatin-sensitive as well as oxaliplatin-resistant cells.     

Combination of chemometrically assisted voltammetry, calorimetry, and circular dichroism as a new method for the study of bioinorganic substances: application to selenocystine metal complexes

Selenium-containing compounds play an important role in antioxidant defense systems, binding to toxic metals, preventing their uptake into cells, and thus protecting cells from metal-induced formation of reactive oxygen species. Here, we present a proposal for a relatively new method as a complement to the more usual methods used in selenium studies. A systematic study of the metal-binding properties of selenocystine (SeCyst) in the presence of divalent metal cations (Cd, Co, Hg, Ni, and Zn) is reported. Isothermal titration calorimetry provides thermodynamic parameters of the systems. Titrations produced curves that could be fit reasonably well to the one set of sites model. The data clearly demonstrate that one M²⁺ binds one SeCyst molecule, and the stable M(SeCyst) complex is formed under these conditions. The order of the SeCyst binding constant for the metal ions is Hg²⁺ > Cd²⁺ ~ Zn²⁺ > Ni²⁺> Co²⁺. Cadmium ion was selected as a modulator for the behavior of SeCyst in the presence of a nonessential metal, and zinc was selected for the case of an essential element. These interactions of SeCyst with Cd²⁺ and Zn²⁺, either individually or combined, were studied in aqueous buffered solutions at physiological pH by differential pulse polarography and circular dichroism spectroscopy. Furthermore, recently developed chemometric tools were applied to differential pulse polarography data obtained in mixtures of SeCyst and glutathione in the presence of Cd²⁺ at physiological pH.     

The alkenyl migration mechanism catalyzed by extradiol dioxygenases: a hybrid DFT study

6-Hydroxymethyl-6-methylcyclohexa-2,4-dienone is a mechanistic probe which when incubated with an extradiol dioxygenase yields a 2-tropolone product. This observation was originally interpreted as evidence supporting a direct heterolytic 1,2-alkenyl migration mechanism for a ring expansion reaction catalyzed by this class of Fe(II)-dependent nonheme enzymes (Xin and Bugg in J Am Chem Soc 130:10422-10430, 2008). In the work reported in this contribution we used quantum chemical methods to test whether such a mechanism is energetically possible and we found that it is not, neither for the mechanistic probe nor for the native catalytic cycle intermediate. Models of increasing complexity were used to calculate energy barriers to the heterolytic 1,2-alkenyl migration and alternative radical mechanisms. It was found that the former involves substantially higher barriers than the latter. A tentative radical mechanism that accounts for the transformation of the probe substrate to 2-tropolone was also proposed, and it involves acceptable barriers.