MTT法药敏试验应用于晚期乳腺癌

[目的]探讨MTT法体外药敏试验在复发转移性乳腺癌化疗中的价值。[方法]取40例复发转移性乳腺癌患者肿瘤组织，以简易机械法制备细胞悬液，应用MTT法体外药敏试验测定其对10种常用化疗药物的敏感性，并指导临床用药，同时与临床疗效进行相关分析。[结果]MTT法检测复发转移乳腺癌患者药敏试验的成功率为92．5％，敏感性为87．5％，特异性为69．2％。阳性预测值为84.0%,阴法预测值为75％，总的预测准确率为81．1％。[结论]MTT简便快速，可作为临床上指导复发转移乳腺癌个体化化疗的有效方法．     

抗体夹心酶联免疫吸附法测定重组E.coli L-门冬酰胺酶研究

用重组E.coliL-门冬酰胺酶免疫家兔，DEAE-纤维素柱层析纯化IgG，采用二步戊二醛交联法将辣根过氧化物酶标记抗体，建立抗体夹心酶联免疫吸附法，测定大鼠血浆中重组E.coliL-门冬酰胺酶浓度。本测定方法的线性范围为1～64U/L，灵敏度为0.4U/L。建立的抗体夹心酶联免疫吸附法测定大鼠血浆中重组E.coliL-门冬酰胺酶具有良好的精密度、回收率、灵敏度和特异性。     

高效液相色谱法测定维生素E烟酸酯胶囊的含量

目的:采用高效液相色谱法测定维生素E烟酸酯胶囊的含量。方法:以Phenomenex Luna C18(250 mm×4.6 mm,5 μm)柱为分析柱,甲醇-0.01％三氟醋酸乙腈(80:20)为流动相,流速1.5 mL·min～1,检测波长264 nm。结果:维生素E烟酸酯在84-839 μg·mL-1范围内峰面积与浓度线性关系良好(r=1.000);平均回收率为100.4％,RSD<1％。结论:本方法准确、简便、快速,适用于测定维生素E烟酸酯胶囊的含量。     

那法瑞林长效制剂对大鼠垂体功能影响

 目的:观察国产那法瑞林(nafarelin )长效制剂的长效作用及对大鼠垂体功能影响，并探讨其作用部位。方法:采用大鼠间质细胞睾酮 (RICT)法检测血清LH(luteinizing hormone)生物活性水平。结果:那法瑞林长效制剂在体内的持续释药时间为((40.0±6.8)d。单次理植3周后大鼠血LH的周期性分泌高峰及激发高峰均消失。预先理植长效制剂组大鼠血LH在切除双侧卵巢后变化不明显。结论:那法瑞林长效制剂确有长效作用，其通过作用于垂体而抑制大鼠垂体功能。     

HPLC法测定清热解毒口服液黄芩苷含量

目的：测定清热解毒口服液中黄芩苷的含量。方法：采用HPLC法，用C18柱，甲醇-水-磷酸（50：50：0．3）为流动相，检测波长276nm。结果：在4．775～191μg/ml的浓度范围具有良好的线形关系。R=1．000（n=6），平均回收率105．6％，日内精密度为1．63％（n=5）。结论：此方法快速、专属，准确。     

Epitopes of human brain acetylcholinesterase

The main purpose of the present work was to identify B-cell epitopes on human brain acetylcholinesterase (AChE) by the synthetic peptide approach. Five hundred and seventy-four decapeptides comprising amino acids No. n to n+9 (where n denotes the residue number of the 583 amino acids in the primary structure of human brain AChE and is an integer in the range 1-574) were synthesized, using the multipin combinatorial chemical synthesis technique, and biotinylated. Epitopes of human brain AChE were detected by enzyme-linked immunosorbent assay (ELISA) and compared with the predicted epitopes of human AChE by 'Goldkey' software. Among 574 synthetic decapeptides, 47 decapeptides at 11 antigenic regions showed immunoreactivity with mouse anti-human brain AChE polyclonal antibodies. The minimum sequence of epitope was defined at every antigenic region explored. The locations and sequences of the former ten continuous epitopes at the 11 antigenic regions of the human brain AChE had been identified as follows: TPVLVWIY (112-119), RTVLVSMNY (143-151), LLDQRLALQW (173-182), RRATQLAH (246-253), VFRFSFVPV (294 approximately 302), KDEGSYFLVY (332-341), RVYA (424-427), LMRY (476-479), KAPQWPPY (496-503), GLRAQACAFW (523-532). The rate of hits of the predicted epitopes from the software came out at 33%. In our work, the epitopes of human AChE have been mapped by purified polyclonal antibody at eleven distinct sites in the primary structure.     

芳香化酶P450在子宫内膜异位症中的表达及意义

目的：探讨芳香化酶P450在子宫内膜异位症发病中的作用。其阳性表达用于诊断子宫内膜异位症可行性。方法：应用免疫组化方法检测正常子宫内膜和子宫内膜异位症的在位及异位内膜芳香化酶P450的表达。结果：芳香化酶P450在正常子宫内膜无表达或弱阳性，在子宫内膜异位症的在位和异位内膜均有强表达，且增殖期高于分泌期表达，敏感度为92％，特异度为95．5％。结论：子宫内膜芳香化酶分泌增加促进子宫内膜异位症的发生，并对其诊断有重要意义。     

海洋岩虫抗菌肽筛选及抗癌活性的初步研究

将岩虫匀浆液经反相浓缩、凝胶过滤和亲和柱层析获得活性组分，在体外用MTS／PMS(氮兰四唑盐／吩嗪硫酸甲酯)方法筛选抗菌肽，再分别用MTS／PMS方法和ELISA—AFP(甲胎蛋白)方法检测其对人肝癌细胞株HepG2增殖、AFP分泌及对正常大鼠成骨细胞株MC3T3-E1增殖的影响。结果表明，纯化的岩虫抗菌肽为组成型碱性蛋白(MW8．1kDa，pI8．6)，不同浓度抗菌肽对肝癌细胞增殖和AFP的分泌均有抑制作用，抑制作用大小与抗菌肽浓度呈正相关性；对正常成骨细胞未见明显的抑制和杀伤作用。     

A continuous fluorometric assay for phospholipase A(2) activity in brain cytosol

Alterations in phospholipase A₂ (PLA₂) activity have been implicated in Alzheimer disease and other neurological disorders, although brain PLA₂ activity is currently measured using lengthy, non-continuous assays. We describe herein a rapid, continuous assay in which we measured PLA₂ activity in mouse brain cytosol (CB-57). Brains were homogenized in HEPES buffer (pH 7.5) and the cytosolic fraction was prepared by centrifugation at 25 000×g for 20 min, followed by centrifugation of the supernatant at 100 000×g for 60 min. Cytosolic protein content was determined using the Bradford assay. Pyrene labeled phosphatidylcholine was added to 50 μg of cytosolic protein in Tris buffer (pH 8.0) containing fatty acid free-bovine serum albumin for a final assay volume of 2 ml. Assay temperature was maintained at 30±1°C. The excitation wavelength was 345 nm and emission was measured at 377 nm. Fluorescence intensity was converted to molar concentrations using a standard curve. Under these conditions, bromoenol lactone inhibited up to 58% of the PLA₂ activity with an IC₅₀ of 0.5 μM. In a separate experiment, lack of appreciable alternative acylhydrolase activity was verified chromatographically. Using this method, brain PLA₂ activity can be measured in a continuous, rapid, and sensitive manner.     

应用微柱凝胶法鉴定婴儿ABO及Rh（D）血型

目的 :比较传统试管法和微柱凝胶法全自动血型系统鉴定婴儿ABO及Rh(D)血型的研究。方法 :选取500份婴儿脐带血标本 ,采用试管法和微柱凝胶法进行ABO和Rh(D)血型 ,并选项取40份抗原性减弱的标本采用两种方法比较。结果 :在鉴定婴儿ABO和Rh(D)血型时两种方法差异无显著性。结论 :微柱凝胶法可取代试管法用于婴儿的ABO和Rh(D)血型鉴定。