人肺癌相关抗原的基因克隆

目的克隆人肺癌相关抗原的基因。方法用λｇｔ１１Ｓｆｉ－Ｎｏｔ定向克隆载体构建人肺鳞癌细胞系Ｌ－７８的ｃＤＮＡ文库,并以抗人肺癌单抗ＡＬＴ－０４为探针进行了筛选。对名为ｈｌｃ－１４的ｃＤ－ＮＡ克隆作了测序分析与进一步研究。用重组的ｐＸＪ４１－ｈｌｃ１４质粒转染ＮＩＨ３Ｔ３细胞,通过免疫组化法对克隆基因进行了表达检测与鉴定,还观察了转染细胞的软琼脂集落形成能力。结果建成１．４×１０６ｐｆｕ／ｍｌ人肺癌ｃＤＮＡ文库,经过四轮免疫学筛选获得６个ｃＤＮＡ克隆,最大的ｃＤＮＡ克隆为ｈｌｃ－１４,长９５４ｂｐ。对该克隆的测序结果与基因库资料比较,未发现同源序列。ｈｌｃ－１４ｃＤＮＡ转染的ＮＩＨ３Ｔ３细胞有明显的肺癌相关抗原表达,并且在软琼脂中的集落形成能力增强。结论ｈｌｃ－１４是一个新的肺癌相关抗原的ｃＤＮＡ序列。这一序列的发现为阐明肺癌的发病机理、探索肺癌基因治疗的靶基因提供了新的线索     

Lead in blood and tissues of mice after administration of low lead doses

Lead levels in whole blood could be determined reliably up to a lower limit of 2 g/100 ml blood, using a modified micromethod of the graphite tube furnace technique. Lead contents of various tissues were also determined by using the automated graphite tube furnace after wet ashing of the organs with nitric acid in autoclaves.Animal experiments with mice showed no measurable increase in blood lead level after a single, 10- or 30-days oral administration of lead in doses of 10–1000 g lead acetate/kg body weight/day. However, these doses led to a rise in tissue lead content. There was a clear dependence of tissue lead content on type of organ examined, lead dose and duration of lead exposure.According to our experiments, the threshold dose which leads to a long-term increase in tissue lead content is assumed to be about 100 g lead acetate/kg body weight/day, orally administered.We are thankful to Prof. Dr. H. Rüssel, Hannover and Dr. M. Fleischer, Saarbrücken for helping in comparison studies.We would also like to thank Mr. H. Dick and Miss. Ch. Hecker for the technical and laboratory assistance.     

Characterization of soluble platelet guanylyl cyclase with peptide antibodies

Summary Soluble guanylyl cyclase partially purified from bovine and human platelets was characterized with antibodies raised against synthetic peptides corresponding to different sequences of the ₁- and ₁-subunits of the bovine lung enzyme. On immunoblots, the platelet guanylyl cyclase was recognized by the four antisera used, with the exception of an antiserum against the C-terminus of the ₁-subunit which did not react with the human platelet but with the bovine platelet ₁-subunit. Furthermore the human platelet ₁-subunit exhibited a slightly lower molecular mass than the bovine protein. The C-terminal antibodies precipitated native platelet and lung guanylyl cyclase activity. In contrast an antibody against a peptide out of the putative catalytic domain, which is highly conserved between all guanylyl cyclases sequenced so far, did not precipitate native guanylyl cyclase, although it recognized both subunits on immunoblots, suggesting that the respective amino acid sequence is located in an inner site of the protein.Abbreviations GC_pep2 YGPEVWEDIKKEA (one letter code) - GC_pep3 SRKNTGTEETEQDEN - GC_pep5 VYKVETVGDKYMTVSGLP - GC_pep8 KKDVEEANANFLGKASGID - TBS-T Tris-buffered saline, containing 0.0501o Tween 20 Correspondence to E. Böhme at the above address     

反相高效液相色谱法测定塞来昔布片含量

目的 :建立反相高效液相色谱法测定塞来昔布片的含量。方法 :采用ZorbaxSB -C18色谱柱 ,以 0 .0 2mol·L-1磷酸二氢钾缓冲液 (pH5 .8) 乙腈 (4 5 :5 5 )为流动相 ,流速为 1.0mL·min-1,以 5 甲基 2 硝基苯酚为内标物 ,检测波长为 2 5 2nm。结果 :塞来昔布在 10 .2～ 5 0 .1mg·L-1范围内呈良好线性 (r =0 .9998) ,平均回收率为 99.8% ,RSD为 0 .35 %。结论 :本法可用于该片的含量测定 ,操作简便 ,结果准确。     

尿石通颗粒中总黄酮的含量测定

目的:测定尿石通颗粒中总黄酮的含量。方法:用70％乙醇提取并制成供试品,采用分光光度法测定。结果:平均回收率为98.28％,RSD=2.13％。3批尿石通颗粒样品中总黄酮的含量分别为1.448,1.403,1.427 mg·g~(-1)。结论:本方法简便、准确、灵敏度高,可作为该制剂的质量控制方法。     

高效液相色谱法测定魔芋精粉中葡甘露聚糖的含量及其单糖组成

目的:建立魔芋精粉中葡甘露聚糖(Konjac glucomannan,KGM)的含量及其单糖组成分析的测定方法。方法:还原单糖用1-苯基-3-甲基-5-吡啉唑酮衍生化后采用反相高效液相色谱法,分段梯度洗脱,检测波长250 nm。通过内标法和外标法的结合对2种单糖的定量进行了方法学研究,同时测定了魔芋精粉中KGM含量。结果:该方法甘露糖在1.001～8.008nmoL线性关系良好(r=0.9994);葡萄糖在1.002～8.016nmoL线性关系良好(r=0.9990),KGM平均回收率为98.1％,相对标准偏差为1.84％,重现性为1.72％,KGM含量为79.5％,其单糖组成为Glc:Man=1:1.51。结论:此方法简便、快速,可同时测定KGM含量及其单糖组成,便于魔芋精粉的质量评价。     

自制中药粉剂中盐酸小檗碱的定量检测方法

目的测定自制中药粉剂中盐酸小檗碱的含量。方法采用薄层扫描法测定了盐酸小檗碱的含量。结果盐酸小檗碱在1～6μg间呈良好的线性关系,r=0.9975,平均回收率为97.95%,RSD为2.36%(n=5)。结论所建立的方法可靠准确地进行定量检测,可用于该制剂的质量控制。     

复方替硝唑含漱液的制备与含量测定

目的:制备复方替硝唑含漱液,建立其含量测定方法并进行稳定性研究.方法:采用最大吸收波长法测定替硝唑的含量,用双波长等吸收点法测定醋酸氯己定的含量,恒温加速试验法考察稳定性.结果:替硝唑回归方程为C＝－0.039 68＋27.491 A,r＝0.999 9,醋酸氯己定回归方程为C＝0.062 47＋50.454 8△A,r＝0.999 99,精密度和回收率良好,置65℃(96 h),70℃(72 h),75℃(48 h)恒温水浴中加热,含量无明显下降.结论:复方替硝唑含嗽液制备方法简单,含量测定方法简便且稳定性好.     

HPLC法测定拉呋替丁及其片剂的含量

目的　建立拉呋替丁及其片剂的高效液相色谱含量测定法。方法　以Shim packODSC18为色谱柱 ,甲醇 0 2mol·L 1醋酸胺溶液 三乙胺 (70∶30∶0 1)为流动相 ,检测波长为 2 75nm。结果　线性系数r =0 9998,平均回收率为 99 6 % (RSD =1 5 % ,n =9)。结论　本方法简便、快速、准确 ,可用于拉呋替丁及其片剂的质量控制。     

人血浆中吡格列酮的固相萃取高效液相色谱法测定

以吡格列酮的同系物 PIOGA为内标 ,采用 C2 固相萃取紫外检测的 HPL C分析法测定人血浆中吡格列酮的浓度。 C1 8分析柱 (15 0× 4.6 mm,5 μm) ,流动相 :水 -乙腈 -冰醋酸 (5 40∶ 46 0∶ 1.2 ,用氨试液调至 p H6 .0 ) ,流速 :1.0 ml/ min,检测波长 2 6 9nm。取血样 0 .5 m l,加入内标经 C2 固相萃取后进样 40μl。最低定量限 (L OQ)为10 ng/ ml,线性范围为 10～ 16 0 0 ng/ ml