聚丙烯酰胺凝胶电泳分析615纯系小鼠血清蛋白

目的：根据各种蛋白质在一定的泳动条件下,都有一定的相对迁移率不同的特点,分析６１５ 纯系小鼠血清中的蛋白质在聚丙烯酰胺凝胶上的分布,进行了定位分析。方法：采用高ｐＨ 不连续性聚丙烯酰胺凝胶电泳及蛋白质特殊染色法。结果：６１５ 鼠血清蛋白质在聚丙烯酰胺凝胶上可分离出１３ ～１５ 条区带,脂蛋白３ 条、白蛋白２ 条、糖蛋白５ 条、铜蓝蛋白１ 条、血红蛋白１ 条、结合珠蛋白３ 条。根据每种蛋白质的Ｒｍ 值定位顺序从阳极到阴极依次为１ ．糖蛋白、２．脂蛋白、３ ．白蛋白、４．脂蛋白、５ ．糖蛋白、６ ．巨球蛋白（ 根据其它资料） 、７ ．铜蓝蛋白、８ ．结合珠蛋白、９ ．Ｈｂ、１０ ．运铁蛋白（ 根据其它资料）、１１ ．脂蛋白、１２ ～１４ ．均为糖蛋白。光密度扫描６１５ 鼠血清蛋白质相对面积比分别为白蛋白为４８－４、α１ 球蛋白为６－９、α２ 球蛋白２３－８ 、β球蛋白为５－０ 、γ球蛋白为１３－８ 。结论：６１５ 鼠血清蛋白质的定位分析对于６１５ 鼠可移植性淋巴细胞型白血病鼠的蛋白质研究具有一定的实际意义,利用Ｒｍ 值来作蛋白质定位分析简单易行,关键在于掌握实验条件的恒定。     

Comparison of the protein and DNA approaches for the characterization of a β-globin chain variant,hemoglobin Cocody [β21 (B3) Asp- → Asn], in a Caucasian patient

Summary Over the past few years, the methodologies used for the identification of hemoglobin A variants have been greatly improved. Both the protein- and DNA-based strategies have their own advantages and limitations. In this report we illustrate the use of both assays for the characterization of a hemoglobin Cocody variant in a woman of Spanish descent. After evaluating the mobility value matrix of the abnormal hemoglobin, the amino acid transition was determined by HPLC and microsequencing of the protein. The ₂₁ Asp was shown to be substituted by an Asn. At the DNA level, the only nucleotide replacement responsible for this amino acid substitution is GAT- AAT at codon 21. The analysis of the-globin gene by denaturing gradient gel electrophoresis (DGGE) method showed that the mutation was situated in a fragment including exon 1. The hemoglobin variant was then identified to be hemoglobin Cocody by DNA sequencing of this fragment.     

Chain length heterogeneity of nucleosomal DNA in mouse liver after dimethylnitrosamine administration

The effect of dimethylnitrosamine on the nucleosomal structure of mouse liver chromatin was studied. After a single oral dose of dimethylnitrosamine (2–75 mg/kg body weight 45 min before sacrifice) liver nuclei were isolated and incubated with micrococcus nuclease. Nucleosomes were separated on sucrose density gradients. There were no differences in nucleosomal sedimentation velocities between preparations from control and dimethylnitrosamine treated animals. The supernatant obtained after centrifugation of the lysed nuclei (2 min at 4,000 g _av) and nucleosomal peak fractions were used for isolation of DNA. DNA was heat denatured in 7 M urea or formamide. After electrophoresis on polyacrylamide gels areas under mononucleosomal DNA and smaller fragments were measured and compared with the total DNA area. The increase in DNA fragmentation was dimethylnitrosamine dose response dependent. When expressed as per cent of controls it amounted to 106% for 2 mg; 115% for 10 mg; 127% for 25 mg; 164% for 75 mg dimethylnitrosamine/kg body weight. A good correlation between mobility and log of chain length of 174 RF DNA-Hae III digest was obtained in nondenaturing 5% polyacrylamide gels and denaturing non-aqueous formamide polyacrylamide gels but not in 12% polyacrylamide gels containing 7 M urea. DNA of mononucleosomal peak fractions contained 200 and that of dinucleosomal peak fractions 400 nucleotides. Fragmentation of DNA was closely related to in vivo dimethylnitrosamine treatment but was not detected in measurements of protein-DNA complexes in the chromatin. It was disclosed on denaturation of DNA followed by polyacrylamide gel electrophoresis.Abbreviations DMN dimethylnitrosamine - SDS sodium dodecyl sulfate The work was supported by Grant Number 1 R0 1 CA26642-01, awarded to A.v.d.D. by the National Cancer Institute, DHEW     

预防非典型肺炎方剂对小鼠外周血液淋巴细胞 DNA损伤的影响

目的　研究预防非典型肺炎方剂对小鼠外周血液淋巴细胞 DNA的损伤作用。方法　应用单细胞凝胶电泳方法测定 6个预防非典型肺炎方剂体内给药对小鼠外周血液淋巴细胞 DNA损伤的作用。结果　ig给予预防非典型肺炎方剂 、 、 (1/ 3临床等效剂量、临床等效剂量、3倍临床等效剂量× 3 d)对小鼠外周血液淋巴细胞 DNA有明显的损伤作用 ,并呈剂量依赖关系。预防非典型肺炎方剂 、 、 (1/ 3临床等效剂量、临床等效剂量、3倍临床等效剂量× 3 d)对小鼠外周血液淋巴细胞 DNA没有明显的损伤作用。结论　预防非典型肺炎方剂 、 、 具有潜在的遗传毒性     

盐酸达克罗宁凝胶的制备

目的：制备盐酸达克罗宁凝胶。方法：用CMC-Na作基质，制备盐酸达克罗宁凝胶，并建立质控方法。结果：处方设计合理，制备工艺可行，平均回收率为100．4％。RSD=O．8％。结论：该凝胶剂质量稳定可靠，可满足临床使用。     

左氧氟沙星滴眼液及眼用凝胶的研究概述

目的 对近年来左氧氟沙星在眼用制剂方面的研究作一综述。方法 参考及整理有关文献。结果 总结并归纳了左氧氯沙星滴眼及眼用凝胶的研究进展。结论：左氧氟沙星缓释型眼用制剂越来越受到重视和发展，并具有广泛的临床应用前景。     

双向电泳技术及其在疫苗研究开发与生产中的应用

双向电泳技术已广泛用于蛋白质组学研究,该技术除了在农业和基础医学领域中有着广泛的应用外,还在疫苗的研究、开发和生产中具有重要的应用前景.此文就双向电泳技术在疫苗抗原及疫苗候选菌株的筛选、基因工程疫苗的研究、抗原呈递及免疫应答机制的研究和疫苗生产过程中批间一致性分析等方面的应用进行了综述.     

菲啰啉对不同氧化剂和多柔比星所致CHL细胞DNA链断裂的影响

目的 为了研究菲啰啉对2种氧化剂和抗癌药多柔比星诱发细胞DNA损伤的影响, 并初步探讨其损伤机制。方法 用不同浓度菲啰啉预处理CHL细胞30 min, 再分别加入3种不同染毒受试物,共同培养一定时间(0.3 mmol·L^-1重铬酸钾：105 min； 0.5 μmol·L^-1多柔比星：5 min； 0.4 mmol·L^-1过氧化氢(H₂O₂)：25 min)后, 用碱性单细胞凝胶电泳方法(ASCGE)测定DNA链断裂情况, 并同时以菲啰啉与二甲亚砜(DMSO，0.33 mol·L^-1)比较对H₂O₂致DNA损伤中·OH的产生和清除。结果 3种染毒受试物均可明显引起CHL细胞DNA链断裂；而当3 μmol·L^-1菲啰啉预处理后, 可使重铬酸钾、H₂O₂所致DNA迁移长度和细胞拖尾率明显降低, 并超过DMSO降低H₂O₂的损伤作用, 当菲啰林浓度升至12 μmol·L^-1时, 可完全消除这两种因素所致的DNA链断裂损伤；10 μmol·L^-1菲啰啉可抑制多柔比星所致DNA损伤, 但浓度直至60 μmol·L^-1仍不能完全消除多柔比星的损伤作用。结论 菲啰啉对2种氧化剂和多柔比星所致DNA损伤均有不同程度的防护作用,同时提示重铬酸钾和H₂O₂所致的DNA损伤主要与需过渡金属离子参与的·OH产生有关, 而多柔比星所致损伤仅部分与此有关。     

Iontophoretic transdermal delivery of buspirone hydrochloride in hairless mouse skin

The transdermal delivery of buspirone hydrochloride across hairless mouse skin and the combined effect of iontophoresis and terpene enhancers were evaluated in vitro using Franz diffusion cells. Iontophoretic delivery was optimized by evaluating the effect of drug concentration, current density, and pH of the vehicle solution. Increasing the current density from 0.05 to 0.1 mA/cm² resulted in doubling of the iontophoretic flux of buspirone hydrochloride, while increasing drug concentration from 1% to 2% had no effect on flux. Using phosphate buffer to adjust the pH of the drug solution decreased the buspirone hydrochloride iontophoretic flux relative to water solutions. Incorporating buspirone hydrochloride into ethanol:water (50:50 vol/vol) based gel formulations using carboxymethylcellulose and hydroxypropylmethylcellulose had no effect on iontophoretic delivery. Incorporation of three terpene enhancers (menthol, cineole, and terpineol) into the gel and when combined with iontophoresis it was possible to deliver 10 mg/cm²/day of buspirone hydrochloride.