人骨骼肌α辅肌动蛋白（α－actinin）的提取及其抗血清的制备和鉴定

本文提取人骨骼肌α辅肌动蛋白（α－ａｃｔｉｎｉｎ）是综合了文献报导有关提取兔肌α－ａｃｔｉｎｉｎ的和提取鸡胗α－ａｃｔｉｎｉｎ的方法,稍加修改而确定的。用Ｈａｓｓｅｌｂａｃｈ－Ｓｃｈｎｅｉｄｅｒ缓冲液提取骨骼肌中的肌球蛋白后,将残余物经硼酸－缓冲液提取、匀浆及高速离心去掉肌动蛋白和肌原纤维的其它成份,上清加硫酸铵至３０％,３５％饱合度所得的沉淀用２２０ｍｍｏｌ／ＬＴｒｉｓ－乙酸溶解、透析、离心后经ＤＥ－５２柱层析可得电泳纯。α－ａｃｔｉｎｉｎ。将人骨骼肌α－ａｃｔｉｎｉｎ纯化制品免疫了三只大耳白纯种家兔,两个多月后,三只兔子都产生免抗人骨骼肌α－ａｃｔｉｎｉｎ的特异抗血清,用双向免疫扩散法和酶联免疫吸附试验（ＥＬＩＳＡ）测定,产生的抗体效价较高,用双扩散法测定效价为１：３２,用ＥＬＩＳＡ测定,用比率法判断结果,效价最高者为１：１００,０００左右,经免疫电镜观察结果证实,上述抗血清可以满足进一步实验要求。     

质粒pcDNA3—HGF的大规模纯经制备研究

质粒ｐｃＤＮＡ３－ＨＧＦ具有潜在的临床治疗缺血性疾病的应用前景。大规模纯化制备是质粒ＤＮＡ应用于基因治疗的关键步骤。质粒大规模纯化制备流程包括：发酵、离心收集细胞、碱性裂解、Ｑ－Ｓｅｐｈａｒｏｓｅ ＸＬ捕获质粒ＤＮＡ、Ｓｏｕｒｃｅ １５Ｑ精制质粒ＤＮＡ,所得纯超螺旋质粒ｐｃＤＮＡ３－ＨＧＦ符合质量标准。该纯化制备方法避免使用动物源性的酶及有毒试剂。     

人骨骼肌α辅肌动蛋白（α—actinin）的提取及其抗血清的制备和鉴定

本文提取人骨骼肌α辅肌动蛋白（α－ａｃｔｉｎｉｎ）是综合了文献报导有关提取兔肌α－ａｃｔｉｎｉｎ的和提取鸡胗α－ａｃｔｉｎｉｎ的方法,稍加修改而确定的。用Ｈａｓｓｅｌｂａｃｈ－Ｓｃｈｎｅｉｄｅｒ缓解液提取骨骼肌中的肌球蛋白后,将残余物经硼酸－缓冲液提取、匀浆及高速离心去掉肌动蛋白和肌原纤维的其它成份,上清加硫酸铵至３０％,３５％饱合度所得的沉淀用２２０ｍｍｏｌ／Ｌ Ｔｒｉｓ－乙酸溶解、透析、离心后     

从睡眠剥夺树qu（TBC）尿液提取内源性“睡眠因子”的研究

本研究报道从睡眠剥夺（ＳＤ）４８－７２ｈ的灵长类原宗Ｔｕｐａｉａ ｂｅｌａｎｇｅｒｉ ｃｈｉｎｅｎ－ｓｉｓ（ＴＢＣ）提取内源性“睡眠因子”Ｓ２Ｃ和Ｓ４Ｂ。收集的尿液经超滤,清液冻干经Ｓｅｐｈａｄｅｘ Ｇ１５分离内源性“睡眠因子”Ｓ２Ｃ和Ｓ４Ｂ。收集的尿液经滤,清液冻干经ＳｅｐｈａｄｅｘＧ１５分离得到Ｆｒａｃｔｉｏｎ Ｉ－Ｖ。活性测定发现Ｆｒａｃｔｉｏｎ－Ⅲ（Ｓ２Ｃ）呈现显著δ－增强促眠效应。经Ｓｅ     

维甲酸对鼻咽癌细胞生长、表型和瘤基因表达的作用

研究了维甲酸（ＲＡ）对鼻咽癌细胞生长、表型和癌基因表达的作用．用ＲＡ诱导鼻咽癌细胞,绘制诱导前后的细胞曲线,观察细胞形态,并用Ｎｏｒｔｈｅｒｎ杂交和ＤＮａｓｅⅠ超敏感区分析法检测基因表达和调控．结果表明,ＲＡ能显著抑制鼻咽癌细胞的生长,前５ｄ下降约５０％．ＲＡ处理后的细胞从典型的多边形形态变成扁平、细长,类似纤维细胞状的形态．ＲＡ诱导前ｃ－ｍｙｃ基因和ｃ－Ｈａ－ｒａｓ基因ＨＮＥ２细胞中高表达,而诱导后ｃ－ｍｙｃ基因表达水平急剧下降,ｃ－Ｈａ－ｒａｓ基因无明显改变．在实验中还发现ＲＡ诱导前后的ｃ－ｍｙｃ基因和ｃ－Ｈａ－ｒａｓ基因中一些重要的超敏感位点和它们的功能．由实验结果可得到如下结论：ＲＡ能促进鼻咽癌细胞分化,通过对染色体上调控位点的作用来抑制ｃ－ｍｙｃ基因的表达,ＤＮａｓｅⅠ超敏感位点与细胞的分化程度、细胞的组织特异性和基因表达状态有关,ｃ－ｍｙｃ基因可通过不同的调控方式而失活．     

两极法与pièce esquillées

两极法与ｐｉèｃｅｅｓｑｕｉｌｌéｅｓ＠陈淳￥复旦大学文物与博物馆学系两极法与ｐｉèｃｅｅｓｑｕｉｌéｅｓ陈淳（复旦大学文物与博物馆学系,上海２００４３３）收稿日期：１９９６－０９－２３１前言两极法或砸击法是最原始和最简单的打片方法之一。它被史前人类甚至现...     

90年代农村发展与农业生态研究

９０年代农村发展与农业生态研究彭廷柏（中国科学院长沙农业现代化研究所,４１０１２５）ＲｕｒａｌＤｅｖｅｌｏｐｍｅｎｔａｎｄＡｇｒｏ－ｅｃｏｌｏｇｉｃａｌＲｅｓｅａｒｃｈｉｎ１９９０ｓ￥．ＰｅｎｇＴｉｎｇｂａｉ（ＣｈａｎｇｓｈａＩｎｓｔｉｔｕｔｅｏｆＡｇｒｉ－ｃｕｌｔｕｒａｌＭｏｄｅｒｎｉｚａｔｉｏｎ,ＡｃａｄｅｍｉａＳｉｎｉｃａ,４１０１２５）．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＥｃｏｌｅｇｙ,１９９３,１２（２）：２－３．ＩｎｏｒｄｅｒｔｏａｐｐｒｏａｃｈｔｈｅｒｅｌａｔｅｄｐｒｏｂｌｅｍｓｏｆＣｈｉｎａ’ｓａｇｒｏ－ｅｃｏｌｏｇｉｃａｌｒｅｓｅａｒｃｈｉｎ１９９０ｓ,ｔｈｉｓｐａｐｅｒａｎａ－ｌｙｚｅｄｔｈｅｎｅｗｔｒｅｎｄｓｏｆｓｔｒｕｃｔｕｒａｌｃｈａｎｇｅｓｉｎｒｕｒａｌｅｃｏｎｏｍｙａｎｄａｇｒｏ－ｅｃｏｌｏｇｙ．ｕｎｄｅｒｃｏｎｄｉｔｉｏｎｏｆｓｏｃｉａｌｉｓｔｍａｒｋｅｔｅｃｏｎｏｍｙｅｘｐｏｕｎｄｅｄｓｏｍｅｖｉｅｗｐｏｉｎｔｓｏｎｔｈｅｃｏｏｒｄｉｎａｔｉｖｅｄｅｖｅｌｏｐｍｅｎｔｏｆａｇｒｏ－ｅｃｏｌｏｇｙａｎｄａｇｒｏ－ｅｃｏｎｏｍｙ,ｔｈｅｓｔｒｅｎｇｔｈｅｎｉｎｇｏｆａｂｓｏｒｂｉ     

L—山梨糖脱氢酶的纯化及性质的研究

从５Ｌ罐发酵Ｌ－山梨糖的Ｇｌｕｃｏｎｉｂａｃｔｅｒ ｏｘｙｄａｎｓ ＳＣＢ３２９和Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ ＳＣＢ９３３混合菌株中差速离心收集ＳＣＢ３２９菌体,破碎,离心获得无细胞抽提液,硫酸铵分级沉淀蛋白后依次经ＤＥＡＥ Ｃｅｌｌｕｌｏｓｅ ５２和Ｑ Ｓｅｐｈａｒｏｓｅ ＦＦ柱层析分得到了Ｌ－册梨糖脱氢酶（ＳＤＨ）,它能将Ｌ－册梨糖脱氢氧化为Ｌ－册梨酮,ＳＤＳ－ＰＡＧＥ电泳测     

福建中亚热带村级农业生态经济类型的成因与发展机制研究

福建中亚热带村级农业生态经济类型的成因与发展机制研究林文雄,吴志强,林群慧,郑世庆,王松良,梁义元（福建农学院,福州３５０００２）ＣｏｎｔｒｉｂｕｔｉｎｇＦａｃｔｏｒａｎｄＤｅｖｅｌｏｐｍｅｎｔＭｅｃｂａｎｉｓｍｏｆＲｕｒａｌＡｇｒｏ－ｅｃｏｌｏｍｉｃＴｙｐｅｓｉｎＭｉｄ－ｓｕｂｔｒｏｐｉｃａｌＲｅｇｉｏｎｏｆＦｕＪｉａｎＰｒｏｖｉｎｃｅ￥．ＬｉｎＷｅｎｘｉｏｎｇ;ＷｕＺｈｉｑｉａｎｇ;ＬｉｎＱｕｎｈｕｉ;ＺｈｅｎｇＳｈｉｑｉｎｇ;ＷａｎｇＳｏｎｇｌｉａｎｇ;ＬｉａｎｇＹｉｙｕａｎ（ＦｕｊｉａｎＡｇｒｉｃｕｌｔｕｒａｌＣｏｌｌｅｇｅ,Ｆｕｚｈｏｕ３５０００２）．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＥｃｏｌｏｇｙ,１９９３,１２（２）：６６－６８．Ｕｓｉｎｇｓｙｓｔｅｍａｔｉｃａｎａｌｙｓｉｓｍｅｔｈｏｄｔｈｅａｕｔｈｏｒｓｓｔｕｄｉｅｄｔｈｅｓｔａｔｕｅｃｈａｒａｃｔｅｒｉｓｔｉｃｓａｎｄｔｈｅｉｒｖａｒｉａｔｉｏｎｐａｔ－ｔｅｒｎｓｏｆｒｕｒａｌａｇｒｏ－ｅｃｏｌｏｍｉｃｔｙｐｅｉｎｍｉｄ－ｓｕｂｔｒｏｐｉｃａｌｒｅｇｉｏｎｏｆＦｕｊｉａｎｐｒｏｖｉｎｃｅ．１１ｔａｐｅｓｏｆｓｙｓｔｅｍｓｔｒｕｃｔｕｒｅａｎ     

鼎湖山土壤的微生物及其对酸度的适应特征

鼎湖山土壤的微生物及其对酸度的适应特征葛荣盛（广东省土壤研究所广州５１０６５０）ＳｏｉｌＭｉｃｒｏｂｃｓａｔＤｉｎｇｈｕｓｈａｎＮａｔｕｒａｌＲｅｓｅｒｖｅａｎｄＴｈｅｉｒＡｄａｐｔａｂｉｌｉｔｙｔｏＡｃｉｄｉｔｙ￥．ＧｅＲｏｎｇｓｈｅｎｇ（Ｇｕａｎｇ－ｄｏｎｇＩｎｓｔｉｔｕｔｅｏｆＳｏｉｌＳｃｉｅｎｃｅｓ,Ｇｕａｎｇｚｈｏｕ５１０６５０）．ＣｈｉｎｅｓｅＪｏｕｒｎａｌｏｆＥｃｏｌｏｇｙ,１９９３,１２（３）：１１－１８．ＴｈｉｓｐａｐｅｒｄｉｓｃｕｓｓｅｓｔｈｅｍｉｃｒｏｆｌｏｒａｅｉｎｓｏｉｌｓｕｎｄｅｒｄｉｆｆｅｒｅｎｔｆｏｒｅｓｔｔｙｐｅｓａｔＤｉｎｇｈｕｓａｎｎａｔｕｒａｌｒｅｓｅｒｖｅａｎｄｔｈｅｉｒａｄａｐｔａｂｉｌｉｔｙｔｏａｃｉｄｉｔｙ：１．Ｕｎｄｅｒｍｉｘｅｄｆｏｒｅｓｔ,ａｍｍｏｎｉｆｙｉｎｇＢａｃｔｅｒｉａ,ｃｅｌｌｕｌｏｓｅ－ｄｅｃｏｍｐｏｓｉｎｇｂａｃｔｅｒｉａａｎｄＯｌｉｇｏｎｉｔｒｏｐｈｉｌｅｓａｒｅｄｏｍｉｎａｎｔ;ｕｎｄｅｒｂｒｏａｄｌｅａｖｅｄｆｏｒｅｓｔ,ｃｅＩｌｕｌｏｓｅ－ｄｅｃｏｍｐｏｓｉｎｇｆｕｎｇｉａｒｅｍｏｒｅ;ａｎｄｌｉｔｔｌｅｄｉｆｆｅｒｅｎｃｅｃａｎｂ     

pH dependence of H+ conduction through the membrane moiety of the H+-ATPase (F0 . F1) and effects of tyrosyl residue modification

A convenient and reliable method to measure passive H+-translocating activity (H+ conductivity) was developed; vesicles reconstituted from the membrane moiety (F0) of H+-ATPase (F0 . F1) and soybean phospholipids were loaded with KCl by a freeze-thaw-sonication procedure and the rate of H+ uptake caused by the K+ diffusion potential upon addition of valinomycin was followed with a pH meter. Of the methods tested, a dialysis method using cholate plus deoxycholate gave the best results for reconstitution. Using this method, H+ conductivity of the membrane moiety of H+-ATPase from a thermophilic bacterium PS3 (TF0) was analyzed. Dependence of H+ conductivity of TF0 on H+ concentration fitted a Michaelis-Menten equation showing a Vmax of 31.3 microgram ion/min . mg of TF0 and a Km of 0.095 microgram ion/liter. Upon modification of a tyrosyl residue of TF0 with iodine, the Km value shifted to 0.71 microgram ion/liter, while the Vmax remained constant. These results were interpreted as indicating that a single tyrosyl residue in N,N'-dicyclohexylcarbodiimide-binding proteolipid of TF0 plays an important role as an H+ donor in the the rate-limiting step of H+ permeation through TF0. TF1, the catalytic moiety of H+-ATPase from the thermophilic bacterium PS3, blocked H+ conduction through TF0. A 1:1 stoichiometry of TF1 and TF0 was found in ATP-dependent membrane potential generation as well as H+ conduction.     

Spectroscopic characterization of the ATPase of the thermophilic bacterium PS3 and its isolated subunits

The ATPase of the thermophilic bacterium PS3, TF0F1, and its subunits has been isolated and their absorption and fluorescence spectra have been measured. The following results were obtained: The tryptophan content of the subunits was determined spectroscopically. Although tryptophan (Trp) and tyrosine (Tyr) are found in TF1, the fluorescence spectrum of native TF1 and its subunits is dominated by Tyr fluorescence; this is in contrast to other proteins. Among (native) TF1 and its subunits only TF1 and the alpha-subunit show a weak fluorescence of Trp, which is blue-shifted, indicating a location in a strongly hydrophobic environment. TF0 fluorescence is dominated by the strong Trp fluorescence. TF0F1 fluorescence is also dominated by the Trp residues. Additionally, its fluorescence is higher than the sum of the isolated TF0 and TF1, indicating marked changes in the microenvironment of the fluorescing aminoacids upon binding of TF1 to TF0.     

Investigation and comparison of the binding between tolvaptan and pepsin and trypsin: Multi‐spectroscopic approaches and molecular docking

Tolvaptan (TF), a selective arginine vasopressin V2 receptor antagonist, was approved by the Food and Drug Administration in 2009. This study mainly investigated the differences between the binding of TF with pepsin and trypsin by using a series of spectroscopy and molecular modeling methods. Thermodynamic parameters and molecular docking results suggested that the binding of TF to pepsin and trypsin were both spontaneous but driven by different forces. For pepsin, the binding was driven by hydrogen bonds and van der Waals forces; but for trypsin, it was driven by electrostatic forces and hydrophobic forces. The quenching mechanism between TF and pepsin and trypsin was investigated by fluorescence experiments and time‐resolved fluorescence spectroscopy. Synchronous fluorescence and 3‐dimensional fluorescence were used to investigate the micro‐environmental and conformational changes of pepsin and trypsin after the insertion of TF. In addition, activity‐measurement results showed that both the pepsin and trypsin activities increased with increasing TF concentration, which may help to understand the possible effect of TF on the digestion and absorption of nutrients in vivo.