赖型钩端螺旋体重组DNApCX7的特异性鉴定和限制性内切酶谱分析

本实验将赖型钩端螺旋体（简称钩体）ＤＮＡ基因库的克隆ｐＣＸ７制备成￣（３２）Ｐ－重组ＤＮＡ探针，对８个不同血清群的１７株问号状钩体、双曲钩体ＰａｔｏｃⅠ株以及细螺旋体３０５５株ＤＮＡ进行打点杂交；同时用１５种ＤＮＡ片断进行限制性内切酶谱分析。结果表明，该重组ＤＮＡ具有问号状钩体种（Ｓｐｅｃｉｅｓ）特异性，但与不同问号状钩体之间的同源性程度有差别；限制性内切酶谱分析发现ｐＣＸ７重组ＤＮＡ片段长约１．７ｋｂ，具有１个Ｂｇ１Ⅱ识别位点和３个ＢｓｔＢ１识别位点。     

伊那普利治疗糖尿病肾病的临床研究

目的评估伊那普利治疗糖尿病肾病(DN)的临床疗效 ,并探讨其作用机制。方法40例持续微量蛋白尿的非胰岛素依赖型糖尿病(NIDDM)患者随机分为常规治疗组(n=19)和伊那普利治疗组(n=21)。利用 131I -邻碘马尿酸钠测定有效肾血浆流量(ERPF) ;肾小球滤过率(GFR)以内生肌酐清除率表示 ;通过ELISA法测定尿微量蛋白 ,包括白蛋白(ALB)、转铁蛋白(TF)、视黄醇结合蛋白(RBP)和N -乙酰 - β氨基葡萄糖苷酶(NAG)。结果伊那普利治疗组增高的尿ALB、TF、RBP和NAG均显著下降 ;ERPF显著增加 ;增加的滤过分数(FF)显著减低。而常规治疗组这些参数无显著变化。结论伊那普利具有改善DN患者的肾小球血流动力学 ,保护肾小球和肾小管功能的作用     

EFFECTS OF ANGIOTENSINS II AND III ON GLOMERULOTUBULAR BALANCE IN RATS

1. The role of angiotensin as a modulator of proximal glomerulotubular (GT) balance was investigated in anaesthetized rats by examining the relationship between glomerular filtration rate (GFR) and absolute proximal reabsorption (APR) during removal of endogenous angiotensin II (AII) and III (AIII) with enalaprilat (CEI) and then during their subsequent replacement by intravenous infusions. 2. Enalaprilat lowered mean arterial blood pressure (MABP) and increased renal blood flow (RBF), GFR, urine flow rate and sodium excretion. Filtration fraction (FF) was not altered. Absolute proximal reabsorption, derived from fractional lithium clearance, increased by only 48% of the change expected for 'perfect' GT balance. 3. Angiotensin II replacement corrected MABP, GFR and plasma renin level, but reduced RBF and increased FF; APR was decreased and GT balance was restored. Urine flow and sodium excretion remained above control values with AII. 4. Replacement with AIII did not correct the hypotension but completely reversed the renal and renin responses to enalaprilat and restored GT balance without affecting FF. 5. It was concluded that the relation between proximal reabsorption and GFR is considerably modified by the intrarenal angiotensin concentration. The findings are best explained by a direct stimulation of proximal tubular sodium transport by angiotensin at the concentrations existing in anaesthetized rats.     

Analysis of the mechanism of action of bradykinin on human basilar artery in vitro

Summary Bradykinin (BK) initially produced concentration-related relaxations of human basilar artery in vitro. Concentration-effect curves constructed at 2 h intervals to BK over an 8 h period were reproducible. The rank order of potency of three kinins on the human basilar artery was found to be BK > methionyl-lysyl-BK > des-Arg⁹-BK. The B₂-receptor antagonist Thi^5,8 d-Phe⁷-BK but not the B₁-receptor antagonist des-Arg⁹-Leu⁸-BK selectively blocked BK-induced relaxations of the human basilar artery.The relaxant effects of bradykinin and acetylcholine but not papaverine were attenuated after removal of the endothelium or treating the tissues with BW755C. Indomethacin was without effect. Concentration-effect curves to angiotensin I were markedly attenuated by captopril at a concentration which had no effect on BK, angiotensin II or 5-hydroxytryptamine responses. It is concluded that BK induced relaxations of the human basilar artery are mediated via activation of a B₂ receptor and the response is dependent upon the release of a factor present in the endothelium. Angiotensin converting enzyme is present in the human basilar artery and is important for the conversion of angiotensin I to angiotensin II but apparently not for the degradation of BK. It is likely that other kininases are present and active in the tissue. Send offprint requests to E. T. Whalley at the above address     

影响α-淀粉酶分离液澄清度的因素

本文根据凝聚和絮凝原理,研究了无机盐和高分子絮凝剂加入量对枯草杆菌BF 7658α-淀粉酶分离液澄清度的影响。实验结果表明,氯化钙可使α-淀粉酶发酵液的ζ电位降低,分离液的澄清度增加,用氯化钙和磷酸氢二钠来处理发酵液,则其分离液的澄清度较单用氯化钙为大,阴离子型高分子絮凝剂如海藻酸钠加入量对分离液澄清度也有一定影响。实验中还发现,澄清的发酵液在10℃下贮存可保持澄清透明状态。     

大肠杆菌表达耐热DNA聚合酶的纯化和鉴定

目的探讨工程菌表达的耐热ＤＮＡ聚合酶的纯化方法。②方法收集ＩＰＴＧ诱导带有耐热ＤＮＡ聚合酶（ＴＤ聚合酶）基因表达质粒的工程菌株ＤＨ－ＴＤ４，用溶菌酶裂解，６０℃处理后，上清液用硫酸铵沉淀，根据溶解度差异进行粗分级，然后进行离子交换层析细分级，并经聚合酶链式反应（ＰＣＲ扩增）鉴定其活性。③结果纯化的ＴＤ聚合酶具有良好的聚合活性。④结论溶解度差异与离子交换层析是工程菌表达的耐热ＤＮＡ聚合酶纯化的主要步骤。基因工程制备的ＤＮＡ聚合酶可用于ＰＣＲ检测     

酵母菌2号培养最佳条件的选择

成人特发性乳糖酶缺乏症的发生率极高，这些人进食牛奶后会出现胃肠胀气、腹痛、腹泄等对牛奶不耐受的消化不良症状。为解决这一问题，自１９８９年起，国内外不少学者是主张有产替代疗法，即将人工培养制备纯化的乳糖酶直接加入牛奶中以水解其中的乳糖来弥补成人乳糖酶的缺乏。     

Structure-activity relationship of macrocyclic and linear gadolinium chelates: Investigation of transmetallation effect on the zinc-dependent metallopeptidase angiotensin-converting enzyme

Transmetallation between commercially available solutions of gadolinium (Gd) chelates and the zinc (Zn)-dependent angiotensin-converting enzyme (ACE) was investigated. In vitro, the strongest inhibitions were observed for the linear Gd complexes, Gd diethylene-triamine pentaacetic acid (DTPA) bis-methylamide (BMA) (IC₅₀ = .016 ± .006 mmol/1) and Gd-DTPA (IC₅₀ = .350 ± .034 mmol/1). The two macrocycles Gd tetraazacyclododecane tetraacetic acid (DOTA) and Gd-HP-DO3A were similar and 400 times less active than Gd-DTPA-BMA. These effects were mainly due to the presence of free ligand for DTPA and calcium (Ca) chelate in the case of DTPA-BMA because the addition of Zn²⁺ in the same quantities suppresses their inhibitory effects. In vivo, these two solutions of linear Gd chelates significantly inhibited ACE activity (Gd-DTPA: 67 ± 9% versus baseline; and Gd-DTPA-BMA: 73 ± 2% versus baseline at the clinical dose of .1 mmol/kg), whereas no significant effect was observed for the two macrocyclic chelates Gd-DOTA and Gd-HP-DO3A. Formulating the Gd chelate solution with either an excess of free ligand or Ca chelate (to decrease Gd³⁺ release) in the case of linear Gd chelate may have deleterious biologic consequences.     

Bioavailability of 1-deamino-8-D-arginine vasopressin with an enzyme inhibitor (aprotinin) from the small intestine in healthy volunteers

Objective: The bioavailability of an aqueous solution of 1-deamino-8-D-arginine vasopressin (dDAVP), with and without an enzyme inhibitor, was studied in six healthy, male volunteers aged 19–34 years, followed for 8 h after each drug administration. Methods: For i.v. administration the subjects received 4 μg dDAVP. For intestinal administration 500 μg dDAVP was administered directly, in two separate sessions, in the first part of the duodenum via a triple-lumen channel tube. In one session a solution of isotonic polyethylene glycol (PEG) was given as a continuous enteral perfusion. In the other session a solution of PEG and aprotinin was administered enterally at the constant rate of 5 ml⋅min⁻¹ for 4 h. Plasma dDAVP was measured using a specific, sensitive radioimmunoassay and intestinal juice was collected for measurement of lipase, chymotrypsin and pH every 30 min for 5 h. Results: The intestinal chymotrypsin activity was decreased after perfusion of aprotinin while the lipase activity was not modified. After i.v. administration, the half-life of elimination of dDAVP was 1.56 h and plasma clearance 1.24 ml⋅min⋅kg⁻¹. The mean bioavailability after duodenal administration of dDAVP + aprotinin was 0.46% compared with 0.09% after duodenal administration of dDAVP alone. The bioavailability of dDAVP after direct duodenal administration of an aqueous solution was similar to that after swallowing a tablet in a previous study and increased 5 times when given together with a perfusion of an enzyme inhibitor. Received: 27 October 1995/Accepted in revised form: 26 February 1996     

A patient with hepatic granuloma formation and angiotensin-converting enzyme production by granuloma cells during clinical relapse of hepatitis A

Elevation of the serum angiotensin-converting enzyme (sACE) level and hepatic granulomas were found during a clinical relapse in a 22 year old patient with acute viral hepatitis type A (AVH-A). The serum transaminase level and sACE level remained high for more than 6 months. In the biopsied specimen of the liver, fibrous rings of granulomas composed of collagen types I, III, and V were observed. Furthermore, the localization of ACE was visible in the rough endoplasmic reticulum of epithelioid cells of granulomas in the liver under electron microscopy using the indirect immunoperoxidase method. These results suggest that granuloma cells in the liver caused by hepatitis A may be involved in ACE production. In addition, other diseases associated with the presence of granulomas in the liver, such as lymphoma, cytomegalovirus infection, visceral leishmaniasis, and lupoid hepatitis, were ruled out. However, the hepatic granulomas disappeared with the healing of AVH-A. In this regard, the present case is considered to be one of the very few cases of hepatic sarcoidosis.