Administração de cafeína durante a gestação altera parâmetros testiculares na prole de camundongos C57/BL6 na vida adulta

Metabólitos ativos da cafeína apresentam toxicidade, podendo causar efeitos deletérios em muitos órgãos no período fetal pelo consumo materno. O estudo objetivou avaliar o papel da administração de cafeína durante a gestação em vários parâmetros importantes para a função testicular em camundongos adultos C57BL/6. Fêmeas grávidas foram divididas em: grupo controle (C), que receberam injeções de sol. salina (0,9% NaCl) e grupo cafeína (CF), que receberam diariamente injeções subcutâneas de 20 mg / kg de cafeína. Filhotes tiveram livre acesso à ração padrão desde o desmame até três meses de idade, quando foram mortos. O grupo CF apresentou uma redução no consumo alimentar (C = 4,36 0,14; CF = 3,79 0,05/g, p<0,05) e ganho de massa corporal (C = 25,73 0,14; CF = 21,08 0,41/g, p=0,0001). Ainda este grupo, apresentou alterações estruturais nos testículos da prole, como redução no peso relativo (C = 0,078 0,003; CF = 0,063 0,002/g, p=0,002), diâmetro tubular (C = 455,4 2,7; CF = 393,5 3,1/m, p=0,0001), lume tubular (C = 310,6 2,5; CF = 254,8 2,9/m, p=0.0001) e altura do epitélio seminífero (C = 144,8 0,9; CF = 138,7 1,3/m, p=0,0005) observados pela técnica de morfometria. Testosterona sérica avaliada por quimioluminescência foi reduzida (C = 6,6 2,8; CF = 0,5 0,2/ng/ml, p=0.04). Western Blotting foi utilizado para análise de expressão protéica. Houve aumento do receptor de leptina (C = 0,81 0,04; CF = 1,28 0,15/g, p=0,01), do receptor para o hormônio leteinizante (C = 0,92 0,05; CF = 0,74 0,05/g, p=0,01),da aromatase (C = 0,32 0,03; CF = 0,48 0,05/g, p=0,03) e do regulador pró apoptose (C = 0,27 0,02; CF = 0,42 0,03/g, p=0,01) enquanto o receptor para hormônio folículo estimulante (FSHR) (C = 0,76 0,06; CF = 0,56 0,04/g, p=0,02), o receptor para proteína reguladora da esteroidogênese (C = 1,009 0,065; CF = 0,584 0,060/g, p=0,0007), a proteína do fator de crescimento vascular endotelial (C = 0,33 0,08; CF = 0,05 0,01/g, p=0,009), o receptor de androgênio (C = 0,97 0,11; CF = 0,59 0,07/g, p=0,02) e o antígeno nuclear de proliferação celular (C = 0,93 0,11; CF = 0,48 0,07/g, p=0,01) foram reduzidos. Este estudo sugere que o consumo de cafeína durante a gravidez parece ser nocivo à fertilidade de animais machos, caracterizando o fenômeno de programação, o que gera alterações funcionais e estruturais nos testículos da prole adulta.

Programação metabólica do testículo pelo tratamento com injeção de leptina em ratos nos primeiros dias de vida

A leptina tem um papel importante na regulação do sistema reprodutivo além de seu papel principal na regulação do peso corporal e ingestão alimentar. O objetivo deste estudo foi avaliar o efeito da administração de leptina durante o período neonatal na função testicular da prole adulta. Vinte e quatro filhotes de 12 mães foram divididos em 2 grupos: Grupo leptina: injetados com 50 L de leptina (80ng/gPC, subcutânea) nos primeiros 10 dias de vida e Grupo Controle: injetados com o mesmo volume de solução salina. Todos os animais foram sacrificados aos 90 dias de vida. Parâmetros analisados: consumo alimentar, massa corporal, crescimento linear, data de início da puberdade, perfil lipídico, níveis séricos de estradiol e testosterona, expressão gênica (PCR em tempo real) e expressão proteica (Western blot) de ObRa, OBRb, aromatase, AR, ER, morfometria testicular, número, morfologia e viabilidade de espermatozoides. Os dados foram expressos como média erro padrão. A significância estatística foi determinada pelo teste t de Student. A Injeção de leptina levou a uma redução (p≤0,008) no consumo alimentar a partir do dia 26 ao 40 e do dia 70 em diante, enquanto que a massa corporal (p≤0,03) e o crescimento linear (p≤0,05) foram reduzidos do dia 26 até o dia 45. O peso da hipófise (p≤0,0006), hipotálamo (p≤0,01), próstata (p≤0,003), testículo (p≤0,008) e epidídimo (p≤0,004) e bexiga (p≤0,009) foram significativamente reduzidos pela injeção de leptina. A Injeção de leptina adiantou o início da puberdade (C=45,0 0,3; L=41,6 0,3; dias, P≤0,0001). Em relação ao perfil lipídico, a administração de leptina gerou um aumento nos níveis séricos de TG (C= 116,5 15,9; L=172,6 19,7; ng/dL, P≤0,05) e uma redução nos níveis de HDL (C=33 1,5; L = 24 3,5; ng/dL, P≤0,02). Os níveis séricos de testosterona também foram reduzidos pela leptina (C=5,2 1,0; L=1,1 0,3; ng/mL, P≤0,003). Todos os genes avaliados por PCR em tempo real mostraram um aumento na sua expressão: Obra (C=0,32 0,04; L=0,69 0,16; P≤0,04), OBRb (C=0,37 0,06; L=0,71 0,16; P≤0,03), AR (C=0,28 0,02; L=0,71 0,16; P≤0,02), Aromatase (C=0,31 0,04; L=0,53 0,09; P≤0,04), ER-α (C=0,79 0,03; L=0,93 0,03; P≤0,01), ER-β (C=0,29 0,03; L=0,73 0,16; P≤ 0,02). Por outro lado, a expressão proteica de OBR (C=4,4 0,29; L=6,6 0,84; P≤0,05), ER-α (C=0,4 0,02; L=0,6 0,05; P≤0,03) e aromatase (C=0,4 0,03; L=0,5 0,02; P≤0,04) aumentaram, enquanto que a expressão proteica de AR (C=0,16 0,01; L=0,09 0,01; P≤0,009) foi reduzida pela administração de leptina. A análise morfométrica mostrou que a leptina levou a um aumento da área total do túbulo seminífero (C=64,6 3,1; L=56,1 2,1;μm, P≤0,01), aumento da área luminal (C=40,6 2,3; L= 48,7 0,9; μm P≤ 0,004) e na altura do epitélio (C=21,5 1,2; L=24,6 1,1; μm, P≤0,03), enquanto que o comprimento do túbulo seminífero foi reduzido no grupo tratado (C=2200 350; L= 1100 110;cm, P≤0,006). A administração de leptina levou a um aumento no número total de espermatozoides (C=20x107 2x107; L=30x107 5x107;Cls/mL, P≤0,009) e no número de anormalidades (C=40,6 1,9; L=45,8 1,2, P≤0,049). Podemos concluir que a leptina tem um papel importante na morfologia e função testicular. A leptina parece ter efeito direto neste tecido uma vez que a expressão gênica e proteica de OBR, AR, ER e aromatase foram alterados pela administração da leptina.

新生豚鼠高胆红素血症时P50抑制变化的研究

目的:评价新生豚鼠高胆红素血症时P50抑制值[即T/C值(T为实验,C为对照)]的动态变化对神经系统的毒性作用.方法:出生5～7天的新生豚鼠30只,随机分成5组,每组6只.其中,第一组为正常对照组(C),其余4组为实验组(T).5组新生豚鼠均在硫喷妥钠麻醉下行颅骨电极包埋术,待手术麻醉清醒后,分别测各组新生豚鼠的T/C值.检测完毕,分别向其中2组实验组动物腹腔注入胆红素溶液100 μg/g,4 h、8 h后观察;另2组实验组动物腹腔注入胆红素溶液200 μg/g,4 h、8 h后观察.正常对照组动物均向腹腔注入生理盐水0.5 ml.各组动物在观察完行为学变化和T/C值检测后,再迅速处死动物,取脑组织,在光镜、电镜下观察脑组织结构的变化.结果:实验豚鼠在注入胆红素溶液后,除Tlb组变化不明显外(P＞0.05),其余各组豚鼠T/C值的变化差异均有统计学意义(P＜0.01).不同实验组与正常对照组的T/C值的变化差异也均有统计学意义(P＜0.01).结论:P50抑制(T/C)在高胆红素血症不同时段均有明显变化,可以较早期地(在胆红素聚集阶段)预测胆红素对神经系统的毒性作用,为临床预防和评价高胆红素血症对新生儿神经系统损伤提供一种新的方法.

原位聚合制备聚乙烯多相材料的微结构调控

聚合物多相材料的制备作为丰富材料品种，扩展材料用途的重要手段受到了广泛地关注和深入地研究。传统的熔融共混法制备聚合物多相材料时，一般需要加入增容剂来改善各相之间的相容性，从而使多相材料的性能达到预期的目标。但是由于增容剂本身也存在分散状态的问题，所以它的加入对多相材料的性能的影响比较复杂。因此，本论文致力于用原位共聚接枝的方法和粒子破碎的方法解决多相材料的界面结合和相分散问题。首先，采用对分散相进行共聚功能化改性的方法，使分散相与乙烯共聚，聚合过程中原位生成聚乙烯接枝物，这种聚乙烯接枝物能起到增容两相的作用，提高两相界面的粘结性，将这种方法应用到有机聚苯乙烯粒子和无机磁性钴粒子体系中，并分别进行了详细的研究；其次，通过聚苯乙烯载体的结构设计，使聚苯乙烯载体催化剂具有较高并且可控的活性，在较高的活性下，聚苯乙烯载体可以破碎，破碎后的聚苯乙烯均匀地分散到乙烯聚合产物中，并且碎片达到纳米级，用这种方法可以改善多相材料的相分散。 本论文的主要工作和研究结果总结如下： 1、采用悬浮聚合制备了交联聚苯乙烯粒子（c-PS），并且在聚苯乙烯粒子的表面引入了双键；c-PS粒子在乙烯填充聚合时，可以与乙烯共聚，从而制备了表面接枝聚乙烯的聚苯乙烯微球（PS-g-PE）；PS-g-PE微球上的聚乙烯的结晶温度与纯聚乙烯的结晶温度相比提高了6℃，说明聚乙烯与聚苯乙烯间的化学连接促进了PE的结晶；PS-g-PE与PE共混后，聚苯乙烯粒子与聚乙烯基体间的界面粘结增强。 2、采用乳液聚合制备了共聚型和不可共聚型交联聚苯乙烯乳胶粒子；将两种聚苯乙烯粒子用于乙烯填充聚合制备了聚苯乙烯/聚乙烯纳米共混材料，结果发现，共聚型聚苯乙烯/聚乙烯的断面上，两相间的界面模糊，并且拉伸断面上也没有不可共聚聚苯乙烯体系中由于拉应力作用而产生的空穴，超薄切片的透射电镜结果同样说明了可共聚型聚苯乙烯体系中界面粘结性的提高；当共聚型聚苯乙烯乳胶粒子的填充量较大（20 wt%）时，聚乙烯共混材料的凝胶含量比较高，说明有更多的共聚型聚苯乙烯在聚乙烯中充当交联点。总之，共聚型聚苯乙烯的填充量在非常少时（0.1 wt%）就能达到很好的改性效果。 3、采用阴离子共聚制备了两亲性的聚苯乙烯-b-聚-2-乙烯基吡啶嵌段共聚物（PS-b-P2VP）和聚4-（3-丁烯基）苯乙烯-聚苯乙烯-聚2乙烯基吡啶的三嵌段共聚物（PBSt-b-PS-b-P2VP）；两个嵌段共聚物在甲苯中均能自组装形成以PVP为核、PS为壳的胶束；Co2(CO)8在PS-b-PVP和PBSt-b-PS-b-P2VP甲苯胶束中热分解得到了由胶束稳定分散的Co磁流体；无水无氧的钴磁流体与乙烯填充聚合后得到了磁性聚乙烯纳米复合材料；钴纳米粒子在聚乙烯中稳定分散，不会发生聚集；PBSt-b-PS-b-P2VP与乙烯共聚后，纳米粒子与聚乙烯基体的相容性进一步提高，从而解决了金属纳米粒子在聚合物中的分散以及界面增强的问题。 4、采用悬浮聚合制备了三种溶胀能力不同的聚苯乙烯交联粒子，研究了溶胀时间对聚苯乙烯载体溶胀程度的影响，以及溶胀程度对乙烯聚合和产物聚乙烯形态的影响；实验结果发现溶胀程度较大、溶胀能力较强的聚苯乙烯载体的负载量和活性都较高；通过提高载体的溶胀程度可以增加催化剂对乙烯聚合的催化活性，最终使载体充分破碎分散到乙烯聚合产物中，原位形成纳米级聚乙烯共混物；乙烯聚合的动力学研究表明载体的破碎是一个由外向内逐步发生的过程；适当的活性可以控制载体破碎的速度，从而得到颗粒形态较好的聚乙烯产物。

大兴安岭冻土湿地泥炭有机碳矿化对气候变化因子的潜在响应

大兴安岭地区是我国地带性多年冻土和冻土湿地的主要分布区，近30年来，大兴安岭地区整体增温显著，气候变化的幅度加大，加之人类活动的频繁，冻土退化严重，冻土湿地出现了原有湿地萎缩和新生湿地扩张的现象。目前，对大兴安岭多年冻土湿地的研究还非常有限，且定性的研究较多，定量的研究极少，多数研究集中于冻土湿地的分布，冻土与湿地之间的机理探讨及描述多年冻土退化对湿地产生的影响等方面。 本论文通过室内培养实验，分析不同温度和湿度梯度及冻融作用下，大兴安岭不同多年冻土区湿地两层泥炭有机碳的矿化状况。结合回归模型，分析大兴安岭多年冻土湿地泥炭有机碳矿化对不同温度和湿度的响应，探讨在气候预案下，大兴安岭多年冻土湿地对气候变化的潜在响应。获得的主要结论如下： （1）大兴安岭多年冻土湿地存在着碳储层，其不同的冻土湿地区由于自然条件、融深等因素的不同，碳储层的厚度也存在着差异。多年冻土湿地含碳量和含氮量都很高，有机碳含量随剖面深度的增加有降低的趋势，泥炭全氮的含量随剖面深度变化复杂，这与湿地土壤形成的气候条件、微地貌和植被类型等有关。大兴安岭连续多年冻土区泥炭，C/N比要高于不连续多年冻土区湿地，并且有机碳含量与全氮含量存在着很好的耦合关系。 （2）大兴安岭多年冻土湿地泥炭有机碳矿化随温度的升高而升高，在培养温度5-20℃下，总的泥炭有机碳矿化量变化范围为18.55~112.91 mg g-1。虽然连续多年冻土区湿地泥炭有机碳矿化率和矿化量都要高于不连续多年冻土区湿地，但经过温度敏感性系数Q10分析，大兴安岭不连续多年冻土区湿地泥炭矿化对温度的响应更显著。从一元动力学方程分析结果来看，大兴安岭多年冻土湿地泥炭有机碳的矿化对15℃响应更显著。 （3）土壤湿度对大兴安岭多年冻土湿地泥炭有机碳矿化产生一定的影响，泥炭总矿化量出现了先随湿度的增加而增加，达到最适宜值后降低的趋势。从本论文的实验设置来看，大兴安岭多年冻土湿地泥炭有机碳矿化的最适宜湿度为60%WHC。利用二元回归模型很好地反映了湿度对大兴安岭多年冻土湿地泥炭矿化的影响，模型推测大兴安岭连续多年冻土区湿地泥炭有机碳矿化的最优湿度为10-20cm层63%WHC，20-30cm层65%WHC；不连续多年冻土区湿地有机碳矿化的最优湿度为10-20cm层65%WHC，20-30cm层59%WHC。 （4）大兴安岭多年冻土湿地泥炭有机碳矿化受温度和湿度的影响显著，其之间的交互作用同样显著。连续多年冻土区湿地有机碳矿化量要高于不连续多年冻土区湿地，这与其含有更高的有机碳和全氮有关。温度和湿度对泥炭有机碳矿化的影响可以用二元二次回归方程很好的表示（P<0.001），通过回归方程和方差分析，结果表明温度和湿度对大兴安岭多年冻土湿地泥炭有机碳矿化都非常重要。 （5）通过培养实验结果显示，虽然温度仍是影响大兴安岭多年冻土湿地泥炭有机碳矿化的主要因子，但随冻融作用处理次数的增加，冻土湿地泥炭有机碳矿化量和温度敏感性系数Q10值有增加的趋势，这意味着冻融作用对大兴安岭多年冻土湿地泥炭矿化产生了不小的影响。虽然冻融作用对大兴安岭多年冻土湿地的影响并不是很大，但大兴安岭处于寒温带，在气候变暖下，冻融过程的频率将加高，冻融作用对大兴安岭多年冻土湿地的影响不容忽视。 （6）大兴安岭地区近30年气候变化趋势分析表明，年均温增长显著，年降水量变化幅度大。在气候变化下，对于不连续多年冻土区，多年冻土不断的退缩及最终的消失，会使冻土湿地萎缩和消失，原有的典型的贫营养的泥炭藓沼泽湿地可能演化为富营养的苔草沼泽湿地或灌丛沼泽湿地，对于大片连续多年冻土区，冻土湿地的变化更加复杂，出现的湿地类型会更多。通过线性气候预案下的大兴安岭多年冻土湿地泥炭有机碳矿化分析，结果显示大兴安岭多年冻土湿地对气候变化响应显著，特别是对于变湿的环境。气候变化下，大兴安岭多年冻土湿地泥炭存在着潜在的分解，多年冻土湿地与气候变化之间存在着正反馈机制。 目前研究表明，大兴安岭地区对气候变化特别敏感，对大兴安岭冻土湿地的研究既填补了国内研究的空白，又对全球的碳循环研究提供了数据支持，并且加深了对冻土湿地生态过程的了解。

土壤微生物和酶对大气CO2浓度与温度升高的响应

土壤微生物(Soil microbes)是生态系统的重要组成部分，它参与土壤中复杂有机物质的分解和再合成，也参与C、N、S、P等的循环。土壤酶(Soil enzyme)是土壤中具有生物活性的蛋白质，它与微生物一起推动着土壤的生物化学过程，并在树木营养物质的转化中起着重要的作用。鉴于土壤微生物和土壤酶对环境变化的敏感性，它们在CO2浓度和温度升高时的反应将在很大程度上影响森林生态系统的结构和功能。因此，要全面评价大气CO2浓度和温度升高对整个生态系统的影响，有必要对CO2浓度和温度升高条件下的土壤微生物的反应进行深入的研究与探讨。本文应用自控、封闭、独立的生长室系统，研究了川西亚高山岷江冷杉（Abies faxoniana）根际、非根际土壤微生物数量，红桦（Betula albosinensis）根际微生物数量以及根际、非根际土壤酶活性对大气CO2浓度(环境CO2浓度+350±25μmol·mol-1，EC)和温度(环境温度+2.0±0.5℃，ET)升高及两者同时升高(ECT)的响应。结果表明： 1) EC和ET显著增加岷江冷杉根际微生物数量，但不同微生物种类对EC和ET的反应有所差异。6、8和10月，岷江冷杉根际微生物数量与对照(CK)相比，EC处理的根际细菌数量分别增加了35%、164%和312%，ET处理增加了30%、115%和209%；EC和ET处理对根际放线菌和根际真菌数量影响不显著。ECT处理的根际放线菌数量分别增加了49%、50%和96%，根际真菌数量增加了151%、57%和48%；而ECT对根际细菌数量影响不显著。EC、ET和ECT处理对岷江冷杉土壤微生物总数的根际效应明显，其R/S值分别为1.93、1.37和1.46(CK的R/S值为0.81)。 2) 红桦根际微生物数量对EC、ET和ECT的响应不同。生长季节(5~10月)，高密度的红桦根际细菌数量与CK 相比，EC的根际细菌数量分别增加28%、33%、423%、65%、43%和79%，而低密度的红桦根际细菌数量增加不显著。ET能显著增加根际细菌数量(7~10月)，其中高密度的根际细菌数量分别增加了377%、107%、35%、22%，而低密度的根际细菌数量分别增加了27%、27%、64%、48%；ECT对两个密度水平下根际细菌数量均未产生有显著的影响。高、低密度的红桦根际放线菌和根际真菌数量与 CK 相比，EC显著增加了低密度的红桦根际放线菌数量，而对高密度的根际放线菌数量无显著影响；ET和ECT对高低密度的红桦根际放线菌数量均未产生显著影响。EC和ET对高低密度的根际真菌数量也无显著影响，而ECT却显著增加了高低密度的根际真菌数量。 3) EC、ET和ECT处理的低密度红桦根际微生物(细菌、放线菌和真菌)数量没有显著高于或低于高密度根际微生物数量，表明短期内密度对红桦根际微生物数量不产生影响。 4) 不同种类的氧化还原酶对EC、ET和ECT的响应不同。5~10月，EC的红桦根际过氧化氢酶活性是CK 的1.44、1.06、1.11、1.10、1.12和1.24倍，差异显著(6月除外)；ET和ECT处理根际过氧化氢酶活性无显著增加。EC的红桦根际多酚氧化酶活性比CK显著增加；ET的根际多酚氧化酶活性显著高于CK(8月除外)。ECT的根际多酚氧化酶活性高于CK，差异不显著。EC的根际脱氢酶活性分别增加了46%、40%、133%、48%、17%和26%，差异显著。5~7月，ET和ECT的根际脱氢酶活性高于CK的脱氢酶活性，而8~9月则相反，差异性均不显著。 5) EC、ET和ECT对不同种类的水解酶的影响不同。EC能显著增加红桦根际脲酶活性，5~10月分别增加了29%、42%，、70%、67%、59%和57%。ET和ECT 对根际脲酶活性未产生显著影响。EC显著提高根际转化酶活性，5、6和9月EC的根际转化酶活性分别比CK高51%、42%和40%。5和10月，ET的根际转化酶活性低于CK，而其余月份却高于CK，但均具有显著性差异。ECT的根际转化酶活性与CK的根际转化酶活性有显著性差异(9月除外)，5、6和7月的根际转化酶活性分别提高了94%、198%和67%。 6) 与CK相比，EC、ET和ECT的非根际土壤微生物数量以及非根际土壤酶活性均无显著提高。EC、ET和ECT的过氧化氢酶、脲酶的根际效应明显，而多酚氧化酶和脱氢酶根际效应不明显。EC和ECT的转化酶根际效应明显，而ET的转化酶根际效应不明显。 It is well known that atmospheric CO2 concentration and temperature are increasing as a consequence of human activities. In past decades, considerable efforts had been put into investigating the effects of climate change on processes of forest ecological system. In general, studies had been mainly focused on the effects of elevated atmospheric CO2 on plant physiology and development, litter quality, and soil microorganisms. Studies showed that there was variation in the responses of root development and below-ground processes to climate between different plant communities. Since the concentration of CO2 in soil was much higher (10~50 times) than in the atmosphere, increasing levels of atmospheric CO2 may not directly in fluence below ground processes. Betula albosinensis and Abies faxoniana, as the dominated tree species of subalpine dark coniferous forest in the western Sichuan province, which play an important role in the structure and function of this kind of forest ecosystem. In our study, effects of elevated atmospheric CO2 concentration (350±25μmol·mol-1), increased temperature (2.0±0.5℃) and both of the two on the number of rhizospheric microbe and rhizospheric enzyme activity were studied by the independent and enclosed-top chamber’ system under high-frigid conditions. Responses of rhizospheric bacteria, actinomycetes and fungi number of Betula albosinensis and Abies faxoniana under different densities(high density with 84 stems·m-2, low density with 28 stems·m-2 ), and rhizospheric enzyme activity of Betula albo-sinensis to elevated CO2 concentration and increased temperature were analyzed and discussed. The results are as the following, 1) In comparion with the control, the numbers of rhizospheric bacteria of Abies faxoniana were increased by 35%, 164% and 312% significantly in June, August and October respectively of EC, and were increased by 30%, 115% and 209% respectively of ET.However the effect of EC and ET on rhizospheric actinomycetes and fungi was not significant. The number of rhizospheric actinomycetes of ECT were increased significantly by 49%, 50% and 96% respectively, and the increment of rhizospheric fungi were 151%, 57% and 48% respectively .The effect of ECT on rhizospheric bacteria was not significant. Rhizospheric effect of soil microbe for all treatments was significant, with the R/S of 1.93, 1.27 and 1.46 for EC, ET and ECT, respectively. 2) Treatment EC improved the number of rhizospheric bacteria of Betula albosinensis under high density significantly in comparison with the control, over the growing season, the greatest increment of rhizospheric bacteria was from July. However, EC had no effect on the number of rhizospheric bacteria under low density. Except May and June, treatment ET improved the number of rhizospheric signifcantly. The effect of treatment ECT on the number of rhizospheric bacteria under different densities was not significant. Of treatment EC, the number of rhizospheric actinomycetes of Betula albosinensis under low density were increased significantly, however, treatment EC did not stimulate the number of rhizospheric actinomycetes under high density. Simultaneously, treatment ET and ECT did not stimulate the number of rhizospheric actinomycetes. Finally, in treatment ECT, the number of rhizospheric fungi under high density were increased significantly, however treatment EC and ET did not stimulate the number of rhizospheric fungi under different densities. 3) Of treatment EC, ET and ECT, the number of rhizospheric microbe of Betula albosinensis under low density were not more or fewer than that of microbe under hign density along the growing season, which showed that plant density had no effect on the nmber of microbe. 4) From May to October, 2004，rhizospheric catalase activity of Betula albosinensis of treatment EC was 1.44, 1.06, 1.11, 1.10, 1.12 and 1.24 times as treatment CK respectively, and the difference was statistically significant(except June). Treatment ET and ECT did not increase rhizospheric catalase activity significantly. In treatment EC, the rhizospheric pohyphenol oxidase activity was higher than treatment CK significantly. The rhizospheric pohyphenol oxidase activity of treatment ET was higher than CK significantly (except August). The rhizospheric pohyphenol oxidase activity of treatment ECT was higher than CK, but the difference was not statistically significant. Over the growing period, the rhizospheric dehydrogenase activity were increased 46%, 40%, 133%, 48%, 17% and 26% respectively by treatment EC, and the difference was statistically significant. From May to July, the rhizospheric dehydrogenase activity in treatment ET and ECT was higher than CK, but from August to October, the rhizospheric dehydrogenase activity was lower than CK, the difference was not significant. 5) Treatment EC increased rhizospheric urease activity significantly, from May to October, rhizospheric urease activity were increased 29%, 42%, 70%, 67%, 59% and 57% respectively by EC. Treatment ET and ECT had no effect on rhizospheric urease activity. Treatment EC improved rhizospheric invertase activity significantly, in May, June and September, the rhizospheric invertase activity of treatment EC were increased 51%, 42% and 40% in comparison with the control. Except May and October, the rhizospheric invertase activity of treatment ET was markly higher than CK. The rhizospheric invertase activity of treatment ECT was significantly different from CK (except September), in May, June and July treatment ECT increased rhizospheric invertase activity by 94%, 198% and 67% respectively. 6) In comparison with the control, treatment EC, ET, and ECT had no effect on the number of non-rhizospheric microbe and non-rhizospheric enzyme activity. Rhizospheric effect of catalase and urease for all treatments was significant, but rhizospheric effect of pohyphenol oxidase and dehydrogenase was not significant. Rhizospheric effect of invertase of EC and ECT was significant, but rhizospheric effect of invertase of ET was not significant.

ACHT操作引起黑麦(Secale cereale)染色体的畸变和重建

用ACHT处理黑麦萌动种子，对修复前后材料的观察和分析结果表明：1. ACHT操作引起染色体数目变化和染色体断裂损失。在一定 条件和范围内，不同处理引起的这种变化具有显著差异，条件越剧烈，染色体数目变化的范围和频率愈大，断片发生的数量和频率 也愈高，同时修复前后染色体数目的变化范围和频率与断片发生的数量和频率以及它们的修复频率均表现明显的相关性。2. ACHT 操作引起染色体畸变的多样性。经ACHT处理后，胚根细胞染色体有4种断裂方式，包括着丝粒断裂、次溢痕断裂、长臂断裂和短臂 断裂等，其中着丝断裂频率最高；产生6种断片类型，包括有着丝粒和端粒的、有着丝粒而无端粒的、有部分着丝粒和端粒的、有 部分着丝粒而无端粒的、只有端粒的、既无着丝粒也无端粒的断片等。3. ACHT操作引起遗传结构重建的多样性。经ACHT处理后， 对修复24-72小时材料进行核型比较(按Stebbins 和 Levan 标准)和随体分析，处理细胞在染色体数目、大小、形态、位置等方面 均发生显著变化，说明ACHT处理使这些细胞的染色体结构和染色体组型发生了深刻变化。进一步通过Giemsa C— 带分析，观察到 多种重建染色体类型，包括易位型染色体、附加型染色体、无着丝粒染色体、化染色体、增加的m染色体以及某些带型特异的染色 体等。4. RAPD 分析从分子水平上验证了ACHT能有效地引起遗传结构的改变。所用10种引物对处理和对照材料基因组DNA的扩增产 物在条带数目、条带位置及带型特征等方面均有明显差异，其中4种引物出现条带减少，6种引物出现条带增加，后者还包括一个带 位移动。这说明两种材料的基因组DNA具有明显的RAPD反应多态性差异。This paper descripes some results draw on the basis of the observation and analysis on the rye before and after repaired through treating its budding seeds by ACHT in contrast to without ACHT: 1. ACHT manipulation caused the number variation and breakage damage of rye chromosome. Within certain conditions and timits, this phenomenon caused by different treats had signifcant difference: the more the treatment condition is drastie, the more the chageable range and frequence of rye chromosomae number, and so is the produced fragments. Meanwhile, there existed striking relationship among the changeable range and frequence of rye chromosome, the produced number and frequence of fragments and repairing frequence. 2. ACHT manipulafion engendered the diversify of rye chromosomal aberration. Four breakage patterns and six sorts of fragment were observed by watching the chromosome of the rye radicle treated by ACHT, including centric breakage (occuring in the highest frequence), secondary constriction breakage, long arm breakage and short arm breakage to the former, Comprising that with both centromere and telomere, that with centromere and without telomere, that with partial centromere and with telomere, that with partfial ceetromere and without telomere, that only with telomere and that neither with centromere nor with telomere, etc. 3. ACHT manipulation engendered the diversify or rye genetic structs reconstruction. Karureotype analysis(according to Stebbins and Levan) and satellite anaeysis were carried out to rye radicle through 24-72-hour-long recoverage after ACHT manipulation, which showed remarkable change happened on the rye chromosomal number、shape、arm ration and pattern, etc. and also on the satellite number、size、shape and location etc. Those indicated that ACHT manipulation engendered violent changes to rye chromatin structure and chromosome type. Further Giemsa C-banding analysis showed many types of reconstructed chromosome, such as translocation、addition、without centromere、st and other chromosome. 4. RAPD analysis checked the validity of ACHT on changing genetic structure of rye on the level of molecular biology. The treated and recovered rye has different amplifying band pattern by using IO valid arbitary primers selected from 40 comparing with the control.

猪场废水短程硝化反硝化-厌氧氨氧化脱氮研究

畜禽废水是农村水环境污染的主要来源之一，其处理的难点在于脱氮。传统生物脱氮法具有能耗高、需大量外加碳源等缺点，开发低成本、高效率的新型生物脱氮技术具有重要意义。 本研究将短程硝化反硝化和厌氧氨氧化两种脱氮新技术结合，让前者为后者创造去除可降解COD、降低总氮负荷、调整pH、调整氨氮和亚硝酸盐氮浓度比例等进水条件，而后者可在无需外加碳源的条件下进一步脱氮，二者结合可成为高氨氮、低C/N废水脱氮的新途径。 试验以低碳氮比猪场废水为研究对象，首先进行了短程硝化反硝化预处理研究，同时启动并运行调控厌氧氨氧化反应器，最后以经过短程硝化反硝化预处理的猪场废水为进水，进行厌氧氨氧化脱氮考察。实验表明：（1）短程硝化反硝化作为厌氧氨氧化的预处理工序是可行的。猪场废水通过短程硝化反硝化，可以达到基本去除可生化COD、部分脱氮、控制出水氨氮和亚硝酸盐氮浓度之比在1︰1左右、pH在7.5～8.0的目的， COD和总氮平均去除率分别为64.3%、49.1%,出水可达到厌氧氨氧化反应的进水要求。（2）采用模拟废水启动厌氧氨氧化反应器，经过5个月左右的运行调控，反应器启动成功并稳定运行，最高总氮去除率为87.1%，总氮容积去除率最高达到0.14kg/m3.d；整个稳定阶段，氨氮、亚硝酸盐氮、硝酸盐氮的变化量之比为1︰1.21︰0.33。（3）经过短程硝化反硝化预处理的猪场废水厌氧氨氧化脱氮效果稳定，氨氮、亚硝酸盐氮、总氮、COD的平均去除率分别为93.0%、99.4%、84.6%、18.1%，处理效果与模拟废水处理系统相比无明显变化。（4）经过短程硝化反硝化预处理后，猪场废水中残留有机物成分在厌氧氨氧化反应过程中无显著变化，主要为酯类和烷烃类物质；残留有机物对厌氧氨氧化效果无明显影响。（5）采用PCR技术进行特殊功能菌种检测，结果表明模拟废水处理系统和猪场废水处理系统的菌群中均含有厌氧氨氧化菌和好氧硝化菌；通过blast比对，厌氧氨氧化菌扩增序列与未培养的Planctomycetales菌和Candidatus Brocadia fulgida菌16S rRNA部分序列相似性分别为95％、90%。（6）MPN法菌种计数结果显示，模拟废水处理系统和猪场废水处理系统的菌群中均含有硝化细菌、亚硝化细菌和少量反硝化菌，实验条件下的微生物系统是一个厌氧氨氧化菌与好氧硝化菌、反硝化菌共存的系统。 Poultry wastewater is one of the main source of water pollution in rural areas，and nitrogen removal is the most difficult part in treating poultry wastewater. There are some disadvantages in traditional nitrogen removal, such as high energy consumption and more additional organic carbon. It is important to develop ecolomical and efficient technologyies. Shortcut nitricfication/denitrification, as a pretreatment process, was combined with Anammox in this research, so that part of total nitrogen and most degradable COD could be removed by the former, and further nitrogen removal could be implemented by the latter. The combination of the two technologies was a new approach to treat wastewater with high ammonium and low C/N. Piggery wastewater with low C/N was treated in lab-scale experiment. Firstly, shortcut nitrification/denitrification was investigated, and Anammox reactor was started up successfully at the same time. Then piggery wastewater after pretreatment was treated by Anammox. The results showed :(1) It was feasible to take nitrification/denitrification as the pretreatment process of Anammox. By using this process, part of total nitrogen and COD were removed, the ratio of ammonium and nitrite reached around 1︰1 and the pH was about 7.8, which were favorable for Anammox. The average removal percentage of COD and total nitrogen were about 64.3% and 49.1%, respectively. (2) Simulated wastewater was used to start up Anammox reactor. The reactor was started up successfully within 5 months and stable performance was achieved. The highest nitrogen removal reached 87.1% and the biggest volumetric total nitrogen removal rate reached 0.14kg/m3.d. The average ratio of ammonium, nitrite and nitrate was 1:1.21:0.33. (3)Taking the effluent of shortcut nitrification/denitrification as the influent, the nitrogen removal efficiency of Anammox was stable, and the the average removal percentage of ammonium, nitrite, total nitrogen and COD were 93.0%, 99.4% , 84.6% and 18.1%, respectively, which had little difference with that by using simulated wastewater..(4) After pretreatment, the residual organic carbon in piggery wastewater showed no obvious change during the Anammox process, and the main organic compounds were saturated hydrocarbon and ester, which had no obvious negative effect on Anammox process.(5) By PCR technology, the existence of Anammox bacteria was confirmed and the aerobic nitrifying bacteria was found to coexist as well. The result of blast showed that the identities of Anammox bacterium to part of 16S rRNA sequence of uncultured Planctomycetales bacterium and Candidatus Brocadia fulgida bacterium were 95% and 90%, respectively.（6）By MPN method, nitrite oxidizer, ammonium oxidizer and denitrification bacteria were detected in both simulated and piggery wastewater treatment system of Anammox, and the microorganism system was composed of Anammox bacteria，aerobic bacteria and denitrification bacteria together.