用限制性核酸内切酶分析钩端端螺旋体DNA的研究

For the application of restriction endonuclease analysis in typing and identifying leptospira, we selected some serovars and isolates, and analysed preliminarily their DNA with four restriction enzymes, EcoR I, Bgl II, Hha I, and Hind III. The DNA samples were isolated from the reference strains and isolates as follows: Serovar lai 56601, 017 (the virulent strain for PDH model of guinea pig), Serovar autumnalis 56606, Serovar manhao II 67020, and isolates 87112 and 87369. Each 2 micrograms of DNA was digested with 20mu of restriction enzyme at 37 degrees C for 2h and electrophoresed in 0.8% agarose gel. The gels were stained in ethidium bromide and photographed with UV light. In our experiments, apparently different restriction patterns of serovar lai 017 were observed with four restriction enzymes. Serovar lai, serovar autumnalis and serovar manhao II showed different patterns with EcoR I, especially in high molecular regions. We also observed in serovar lai 017 a distinct 10.5kb band which was obscure in 56601, the reference strain of serovar lai, after EcoR I digestion. The three serovars showed some delicate differences in Hind III restriction pattern. The two isolates from Apodemus agrarius in Sichuan (1987) 87112 and 87369 had patterns identical to those of serovar lai 56601, 017 with EcoR I, and 87112 also had a pattern identical to 56601, 017 with Hind III. Our results indicate that selected three serovars can be identified by analysis of their DNA with EcoR I and Hind III. It is suggested that restriction endonuclease analysis be a good method in typing and identify leptospira and in studying the differences of special DNA molecules.     

用限制性核酸内切酶分析钩端螺旋体DNA的研究

作者选用钩体赖型56601株、017株、秋季型56606株、曼耗2型67020株、1987年分离株87112、87369株,用内切酶EcoR Ⅰ、Bgl Ⅱ、Hha Ⅰ和Hind Ⅲ消化分析其DNA,结果表明:钩体017株的四种内切酶图谱明显不同,在EcoR Ⅰ酶切图谱上,赖型、秋季型、曼耗2型钩体DNA的主要差别在高分子区带,赖型强毒力株017较同型弱毒力株56601在10.5kb处染色较深;三型钩体的Hind Ⅲ酶切图谱仅有细小差异;分离株87112和87369的EcoR I 酶切图谱与赖型56601和017基本一致。在HindⅢ酶切图谱上,分离株87112与赖型56601株、017株相似。提示:限制性核酸内切酶分析可以作为钩体分型和鉴定的手段。     

L-NMA的合成及其化学和生物学初步鉴定

一氧化氮(NO)是当前深受重视的一个细胞因子,其合成酶的抑制剂L—甲基精氨酸(L-NMA)是NO研究领域里的一个重要工具药物。本实验室以甲基硫脲和鸟氨酸为原料,成功地合成了L-NMA的盐酸盐。经化学鉴定和仪器分析,证明该产品是目标化台物。生物学鉴定以     

牛磺胆酸钠致大鼠急性出血坏死性胰腺炎早期胰腺血流的测定

作者以5％的牛磺胆酸钠(Na—Tc)注入大鼠胆胰导管内,引起急性出血坏死性胰腺炎,以注射生理盐水为实验对照组,生化测定血清脂肪酶和淀粉酶活性,结果于注射Na—Tc后3小时血清脂舫酶和淀粉酶活性显著升高,12小时最明显;用~(86)RbCl示踪法测定,显示胰腺血流量和胰组织灌流量于注射Na—Tc后1小时明显升高,并维持较高水平至12小时下降。这些结果提示:Na—Tc致大鼠急性出血坏死性胰腺炎早期胰腺血流量增加,为一般炎症充血反应。     

牛磺胆酸钠致大鼠急性出血坏死性胰腺炎胰腺组织的光镜和电镜观察

作者以5％的牛磺胆酸钠(Na-Tc)注入大鼠胆胰导管内引起急性出血坏死性胰腺炎。光镜和电镜观察结果显示:注入Na-Tc后10分钟到1小时,胰腺小叶出现细胞膜性结构溶解的灶性坏死,血管少许破坏和轻度出血;注入Na-Tc后3到 6小时,胰腺多个小叶成片坏死,血管溶解和重度出血。坏死前的胰腺细胞内出现大量含有酶原颗粒等细胞器的自噬泡。     

缺血预处理对心肌细胞及其膜结构的保护作用和机理的实验探讨

作者采用Ｌａｎｇｅｎｄｏｒｆｆ灌流模型，观察了反复短暂心肌缺血（ＲＢＭＩ）对心肌形态学、收缩功能、冠脉流量（ＣＦＲ）、心肌细胞膜磷脂（ＰＬ）含量及５’—核苷酸酶（５’－ＡＭＰａｓｅ）比活性的影响。结果表明：再灌注后，心率、左室内压及ＣＦＲ在两组间无显著性差异，但实验组的ＰＬ含量及５’－ＡＭＰａｓｅ活性均高于对照组，超微结构观察也显示实验组心肌结构保存较对照组好。本结果提示，ＲＢＭＩ对缺血心肌膜结构有明确保护作用，且该作用与ＣＦＲ无关。     

用限制性核酸内切酶分析钩端螺旋体DNA的研究

作者选取钩端螺旋体(简称钩体)赖型017株、赖型56601株、秋季型56606株、曼耗2型67020株;1987年分离的87112株和87369株按本窒方法提取DNA,用BgIⅡ、EcoRI;Hha Ⅰ和Hind Ⅲ20u37C°消化2ug钩体DNA2小时,予0.8％琼脂糖凝胶上100V电泳4小时后照象记录。结果显示:不同内切酶对同一钩体DNA酶切显示不同图谱,EcoRI酶切图谱上,赖型、秋季型和曼耗2型明显不同,主要差别在高分子区带,赖型参考菌株56601与同型强     

牛磺胆酸钠致大鼠包性出血坏死性胰腺炎早期胰腺血流的测定

Acute haemorrhagic necrotizing pancreatitis was induced by injecting 5% sodium taurocholate (Na-Tc) directly into the common biliopancreatic duct in rats. In control group 0.9% NaCl was used. The activity of serum lipase and amylase distinctly increased at 3 h and went up to the maximum at 12 h after injection of Na-Tc. The pancreatic blood flow and tissue perfusion per gram increased apparently at 1 h and decreased at 12 h after injection of Na-Tc by using the fractional indicator distribution technique with 86RbCl. The results demonstrated that the early stage of acute haemorrhagic necrotizing pancreatitis induced by Na-Tc in rats was still a primary inflammatory response.     

牛胆酸钠所致大鼠急性出血性胰腺炎模型及早期病理变化的探讨胰血流、血清脂肪酶和淀粉酶活性的测定及光镜、电镜的观察

作者用5％牛胆酸钠溶液直接注入大鼠胰导管,引起急性出血性胰腺炎,在给药后1小时、3小时、6小时和12小时以Rb~(86)Cl测定胰腺血流量,与生理盐水对照组相比,6小时胰腺血流量增高72％(P<0.01),胰腺组织灌流量增加41％(P<0.05),12小时胰腺血流量和胰腺组织灌流量下降。给药后1小时、3小时和6小时测定血清脂肪酶、淀粉酶活性     

牛磺胆酸钠致大鼠急性出血坏死性胰腺炎早期胰腺血流的测定

作者以5％的牛磺胆酸钠(Na-Tc)注入大鼠胆胰导管内,引起急性出血坏死性胰腺炎,以注射生理盐水为实验对照组,测定血清脂肪酶和淀粉酶活性,结果注射Na-Tc后3小时此两酶的活性均显著升高,12小时最明显;用~(36)RbCl示踪法测定,显示胰腺血流量和胰组织灌流量于注射Na-Tc后1小时明显升高,并维持较高水平至12小时下降。这些结果提示:Na-Tc致大鼠急性出血坏死性胰腺炎早期胰腺血流量增加,为一般炎症充血反应。