三氧化二砷对肝癌细胞系HepG2生长及核基质蛋白的影响

目的 探讨三氧化二砷对人肝癌细胞系ＨｅｐＧ２的生长及核基质蛋白的影响。方法 应用生长指数分析和高分辨ＳＤＳ－聚丙烯酰胺交双向电泳比较用Ａｓ２Ｏ３处理后的ＨｅｐＧ２细胞主核基质蛋白的改变。结果 Ａｓ２Ｏ３抑制ＨｅｐＧ２的生长,双向电泳显示经Ａｓ２Ｏ３处理后ＨｅｐＧ２细胞内出现新的核基质蛋白,并且该蛋白是细胞特异性的。结论 核基质蛋白是某些抗癌药物的作用靶子,Ａｓ２Ｏ３抑帛 癌细胞的生长可望有用于临床     

一种新的新生儿溶血病实验诊断技术—微柱凝胶技术

目的建立一种新的微柱凝胶抗球蛋白实验诊断新生儿溶血病。方法采用微柱凝胶抗球蛋白实验与试管抗球蛋白实验同时平行做三项实验:直接抗球蛋白实验、游离IgG抗体测定、放散实验。结果两法检测126例(Rh(D)均为阳性)疑为新生儿溶血病标本,阳性率分别为76例(60.3％)、50例(39.7％)。结论该法具有操作简单、方便、标本用量少、结果易判定、敏感性高、特异性强、结果可较长时间保存等优点。与临床诊断符合率高达98％,为新生儿溶血病提供实验室可靠诊断依据。     

丙烯腈对大鼠外周血淋巴细胞DNA损伤的研究

［目的］ 探讨丙烯腈对大鼠外周血淋巴细胞ＤＮＡ的损伤作用。［方法］ 在急性和亚急性经口染毒试验中,采用单细胞凝胶电泳技术检测不同染毒剂量（１０ｍ ｇ／ｋｇ、３０ｍ ｇ／ｋｇ 和５０ｍ ｇ／ｋｇ）丙烯腈（ＡＣＮ）及不同染毒时段（２ｈ、１４ｄ、２８ｄ、和４２ｄ）对大鼠淋巴细胞ＤＮＡ 的损伤情况。［结果］ 在急性和亚急性染毒试验中,大鼠淋巴细胞ＤＮＡ损伤程度均随ＡＣＮ 染毒剂量增大或染毒时间延长而逐渐加重,并显著高于对照组或染毒１４ｄ 组（Ｐ＜０．０１）,且存在较好的剂量－反应和时间－反应关系（Ｐ＜ ０．０１）。亚急性染毒试验中,随着染毒剂量的升高和染毒时间的延长,淋巴细胞ＤＮＡ 的损伤程度可达到饱和。染毒组细胞ＤＮＡ 损伤反应模式明显不同于对照组,单个细胞间的差异较大,且这种差异随染毒剂量增大和染毒时间延长也逐渐增大。［结论］ ＡＣＮ对大鼠外周血淋巴细胞ＤＮＡ具有明显的损伤作用,且损伤程度和模式受ＡＣＮ 染毒剂量和染毒时间的共同影响。     

中药复方962对老年大鼠外周血淋巴细胞DNA损伤的影响

目的：观察不同年龄组（4月和24月龄）大鼠和连续灌胃给予中药复方962 3个月的老年大鼠的外周血淋巴细胞DNA的损伤情况。方法：利用快速、灵敏的单细胞电泳技术结合激光扫描共聚焦显微镜来观察大鼠外周血淋巴细胞的DNA损伤,以彗星样细胞出现率（计数一定量细胞中彗星样细胞所占的比例）、占彗星长度（“彗星”电泳方向上的最大长度）和尾距（尾部DNA占总DNA的百分比与头尾部中心间距的乘积）来评价淋巴细胞DNA的损伤程度。结果：发现老年大鼠外周血淋巴细胞中彗尾样细胞出现率显著高于青年大鼠（P＜0．01）,复方962不仅能明显降低老年大鼠的淋巴细胞的彗尾样细胞出现率,而且能明显降低老年大鼠外周血彗星样细胞的总彗星长度和尾蹑（P＜0．01）。结论：实验表明复方962对老年大鼠淋巴细胞DNA的损伤有明显的保护作用。     

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。     

长沙地区汉族人群红细胞EsD和PGM1的分布研究

本文采用琼脂糖、水解淀粉混合凝胶电泳法同时分析血液EsD、PGM1二种酶型。调查了这二种酶的表型分布和基因频率及其识别能力 ,并与不同地区、不同国家和不同民族进行比较研究。此法可用于个人识别和亲权鉴定     

精神分裂症脑脊液中相对分子质量130 ku蛋白质含量增加的免疫学及临床意义

目的 研究精神分裂症脑脊液中存在的特异性ＩｇＧ抗体。方法 应用ＳＯＳ－聚丙烯酰胺凝胶电泳及蛋白印迹技术对精神分裂症脑脊液与对照脑脑液同时进行检查。结果 精神分裂症脑脊液中相对分子质量１３０ｋｕ的蛋白质含量显著增加,在蛋白印迹分析中发现此蛋白质可与羊抗人ＩｇＧ抗体发生免疫结合反应,结论 提示该蛋白质组分可能是一种与ＩｇＧ相关的蛋白质。     

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