Survival, mutagenesis, and host cell reactivation in a Chinese hamster ovary cell ERCC1 knock-out mutant

Positive selection-negative selection gene targeting was usedto disrupt the nucleotide excision repair gene ERCC1 in a Chinesehamster ovary cell line, CHO-K1. Southern and Northern analysisshowed that a cell clone isolated by this targeting approach,CHO-7-27, had an ERCC1 gene structure consistent with targeteddisruption of ERCC1 exon V, and did not express ERCC1 mRNA.CHO-7-27 was further characterized with respect to UV and mitomycinC sensitivities, and was shown to exhibit severe mutagen sensitivityphenotypes consistent with those of other CHO cell ERCC1 mutants.Mutation frequency experiments showed that CHO-7-27 was UV-hypermutableat the hypoxanthine-guanine phosphoribosyltransferase locus.Experiments assessing host cell reactivation of viral DNA synthesisfor UV-irradiated adenovirus showed that CHO7-27 exhibited aseverely deficient HCR phenotype similar to that of UV20 cells.Our results demonstrate that CHOK1 cells are hemizygous forthe ERCC1 gene, and show that the comparatively mild mutagensensitivities and lack of severely deficient HCR phenotypesof conventionally derived CHO-K1 ERCC1 mutants, in contrastto the severe phenotypes of CHO-AA8-derived mutants, are notdue to any intrinsic genetic differences between CHO-K1 andCHO-AA8 parental cell lines. ⁴To whom correspondence should be addressed     

从自由基生物学角度评价鸡肉质量的实验研究

本文把外观健康和非健康的活鸡各20只分成两组,宰后测其肝、腿肌中超氧化物歧化酶(SOD)含量和脂质过氧化产物丙二醛(MDA)水平,发现两组中上述指标值差异显著(P<0.050 ;同时对两组实验鸡胴体在宰后4小时做了感官鉴别,也判断出其品质存在差异。提示体内SOD活性和MDA水平可以反映鸡肉质量,比感官鉴别更具敏感性,同时也可以反映宰前鸡的健康状况。     

天然药物中硒的正交设计石墨炉原子吸收测定

目的:建立天然药物中硒的含量测定方法。方法:应用正交设计法优化石墨炉原子吸收光谱硒的分析条件,特别是混合基体改进剂和石墨炉工作参数的选择。结果:线性范围为5～100ng/ml,相关系数r=0.9994,相对标准偏差RSD<10％,用国家标准物质检验本方法,准确度和精密度良好。结论:本方法适合天然药物中硒的测定     

术前左室射血分数及短缩分数对冠状动脉旁路术后恶性心律失常的预测价值

为了评价冠状动脉旁路术 (CABG)术前左室射血分数 (LVEF)及左室短缩分数 (LVFS)对术后室性心律失常 (VA)预测的准确性 ,采用术前及术后 2周心脏彩超EF、FS值 (面积长轴法 )、心室晚电位 (VLP)、心肌酶、持续心电监测的方法 ,对我院 1 5 0例行CABG术的患者进行分析。结果 :1 )术前心肌梗死 (MI)、室壁瘤、VA及VLP阳性患者EF、FS值明显减低 ;2 )术前左心功能不全 (LVD)患者术后EF、FS值明显改善 ;3 )术前LVD、VA、VLP阳性及室壁瘤患者术后VA发生率明显高于其他患者。提示 :1 )面积长轴法EF、FS值是反映左心功能的敏感指标 ;2 )术前LVD患者术后短期左心功能明显好转 ,获益最大 ;3 )非LVD患者术后因心肌顿抑导致近期心功能暂时下降 ;4 )EF≤ 4 0 %和(或 )FS≤ 2 4 %是预测术后VA的独立指标 ,FS较EF更能准确地反映心脏收缩功能 ;5 )LVD、VLP、室壁瘤等综合指标分析有助于提高对术后VA预期的敏感性、特异性和准确性     

刺五加皂甙对培养神经元拟衰老反应的观察

目的 :研究刺五加皂甙 (ASS)对神经细胞老化的保护作用。方法 :采用无血清培养方式制备离体培养的ICR小鼠胎鼠脑皮层神经元衰老模型 ,从生化和形态方面观察刺五加皂甙对衰老条件下的神经元保护作用。结果 :对照组MTT(2 .782± 0 .0 2 8) ,LDH (2 3.0 7± 2 .6 4 ) ,与正常组比较P <0 .0 1。研究组加用 1.2 5mg/10 0ml,2 .5mg/ 10 0ml和 5mg/ 10 0ml浓度的刺五加皂甙后 ,MTT依次为 (0 .0 82 7± 0 .0 2 3) ,(0 .86 1± 0 .0 2 6 )和(0 .90 5± 0 .0 98)。LDH依次为 (2 0± 1.0 5 ) ,(18.4 1± 1.6 4 )和 (18.5 5± 3.83) ,与对照组比较P <0 .0 1～ 0 .0 5。ASS能明显提高衰老神经细胞的MTT活性 ,降低LDH的活性 ,显微镜及扫描电镜观察 ,加用刺五加皂甙保护的研究组神经细胞损伤明显减轻 ,部分细胞形态基本趋于正常。结论 :刺五加皂甙能明显提高衰老神经细胞的MTT活性 ,降低LDH活性。有效地延缓神经元细胞的衰老。     

正常及良性前列腺增生组织中表皮生长因子活性的测定

为了解表皮生长因子在前列腺组织中的作用，利用免疫组织化学方法和竟争放射受体法对人正常和良性前列腺增生组织中表皮生长因子的组织定位和活性进行了检测。结果表明表皮生长因子主要定位于前列腺的分泌上皮细胞，正常前列腺和良性前列腺增生组织中表皮生长因子的活性无显著性差异。提示前列腺组织中表皮生长因子的主要作用可能并非在于调节前列腺上皮细胞的增殖。     

X线和苏拉明联合应用诱导肾癌GRC—1细胞凋亡的研究

为寻求敏感而有效的治疗肾癌的方法，采用Ｘ线照射和不同剂量的苏拉明联合应用的方法诱导肾癌ＧＲＣ１细胞凋亡，以原位缺口末端标记和ＤＮＡ片断梯度电泳分析法检测。结果显示：本法可诱导肾癌ＧＲＣ１细胞凋亡，并随苏拉明剂量的增加，凋亡细胞增多（最高６８．２％）。随时间延长凋亡细胞数增多（最高达６６．２％）。ＤＮＡ片断梯度电泳分析表明，随苏拉明剂量增加和给药后时间的延长，ＤＮＡ片断梯度改变更加明显。结果提示Ｘ线和苏拉明联合应用治疗肾癌，可望取得良好的治疗效果。     

Root avulsion of brachial plexus in infants and children.

Twenty-one patients, under 6 years of age, with root avulsion of the brachial plexus were treated from 1975 to 1987. Among them, 9 had root avulsion of the upper trunk, 2 root avulsion of the lower trunk with middle trunk broken, and 10 root avulsion of the whole brachial plexus. Follow-up of 16 patients showed excellent results in 2 patients, good in 11, fair in 1 and poor in 2. It is suggested that good results can be obtained if multi-paired nerve transfer is adopted in treatment. We conclude that owing to anatomical and physiological characteristics of the respiratory system in children, it is harmful to perform phrenic nerve transfer concomitantly with intercostal nerve transfer. Advisably, two-stage operation is required, otherwise dyspnea and pneumonia are liable to occur. In addition, the time of the operation could be extended to 4-5 years after injury in children, but best results are obtained if the operation is performed within 1 year.

     

Pulmonary artery balloon counterpulsation for right ventricular failure after right ventriculotomy in the swine

To assess the efficacy of intrapulmonary balloon counterpulsation in the management of right ventricular failure after right ventriculotomy, we undertook an experimental study in a swine model. To mimic the clinical settings more closely, (1) we left the automatic control of the heart intact (2) did not use cardiopulmonary bypass to support the left side of the heart, and (3) induced right ventricular failure by means of a generous surgical incision (50% to 70% of the anterior wall) of the right ventricle. The criteria set for right ventricular failure were (1) 50% increase in right ventricular end-diastolic pressure, (2) 30% decrease in mean arterial pressure, and (3) 30% decrease in cardiac output. Right ventricular failure was attained in all animals studied: A 230% increase in right ventricular end-diastolic pressure, a 43% decrease in cardiac output, and a 34% decrease in mean arterial pressure were evident after the right ventriculotomy. A specially designed intrapulmonary balloon catheter (Datascope Corp., Oakland, N.J.) was placed into the left pulmonary artery through the right ventricular outflow tract. A Datascope console was used for counterpulsation. Effects of counterpulsation for 40 minutes in a 1:1 mode were assessed after surgical induction of right ventricular failure in 14 swine. Each animal served as its own control. The mean hemodynamic changes are outlined: Right ventricular end-diastolic pressure decreased by 48.9% (p = 0.01). Mean arterial pressure increased by 68.8% (p = 0.01) and cardiac output by 44.2% (p = 0.01). Histologic studies disclosed no morphologic damage to the pulmonary artery or valve in the specimens analyzed. In addition, these results were compared with those in a second group of seven swine in which right ventricular failure was induced by right ventriculotomy and a balloon was placed into the left pulmonary artery but not activated. These results of short-term counterpulsation should be evaluated in a longer term model so as to mimic more closely the clinical setting. If the hemodynamic benefits are duplicated, intrapulmonary balloon counterpulsation should be considered as a simple, effective device when right ventricular failure develops after right ventriculotomy. It effectively improves right ventricular function without damaging the pulmonary artery or valve.