伤寒杆菌稳定L型粗糙型返祖菌的蛋白图谱和抗原研究

甲型副伤寒杆菌温和噬菌体转导伤寒杆菌稳定Ｌ型返祖形成的两株粗糙型伤寒杆菌,在ＳＤＳ－ＰＡＧＥ电泳中缺失多条高分子量蛋白带。抗原分析表明,两株粗糙型返祖菌具有同其亲代伤寒杆菌一致的Ｈ抗原,但缺乏Ｏ抗原以及伤寒杆茵和甲型副伤寒杆茵共同的某些表面抗原。     

幼年和成年小白鼠LDH与SOD动态变化的电泳分析

采用聚丙烯酰胺凝胶圆盘电泳分析方法,研究了幼年鼠和成年鼠睾丸乳酸脱氢酶（ＬＤＨ）同工酶与超氧化物歧化酶（ＳＯＤ）的酶带变化。观察结果表明,成年鼠睾丸的凝胶电泳胶条,多数显现六种ＬＤＨ同功酶带,其中ＬＤＨ５和ＬＤＨ４占优势,ＬＤＨ１、ＬＤＨ２、ＬＤＨ３显带较弱,在正极端ＬＤＨ的下面是第６条酶带,相当于Ｃ亚单位（ＬＤＨｃ）的Ｃ４。幼年鼠睾丸的凝胶电泳胶条,只显现ＬＤＨ１和ＬＤＨ２两条同功酶带,以ＬＤＨ     

天然及氧化修饰脂蛋白对人动脉平滑肌细胞原癌基因…

动脉平滑肌细胞（ＳＭＣ）的增殖在动脉粥样硬化（ＡＳ）的形成过程中极其重要。我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣ ｓｉｓ,ｊｕｎ,Ｈ－ｒａｓ原癌基因及Ｒｂ抗癌基因转录表达的影响。结果表明：（１）ＨＤＬ对ＳＭＣｓｉｓ,ｊｕｎ,ｒａｓ基因表达无影响;（２）ＬＤＬ和ＶＬＤＬ有使这些基因表达增加的趋势;（３）ｏｘ－ＬＤＬ,ｏｘ－Ｖ     

天然及氧化修饰脂蛋白对人动脉平滑肌细胞原癌基因表达的影响

动脉平滑肌细胞（ＳＭＣ）的增殖在动脉粥样硬化（ＡＳ）的形成过程中极其重要。我们在建立人主动脉ＳＭＣ体外培养方法的基础上,观察了ＬＤＬ,ＶＬＤＬ及ＨＤＬ和相应的氧化修饰型脂蛋白对培养人ＳＭＣｓｉｓ,ｊｕｎ,Ｈ－ｒａｓ原癌基因及Ｒｂ抗癌基因转录表达的影响。结果表明：（１）ＨＤＬ对ＳＭＣｓｉｓ,ｊｕｎ,ｒａｓ基因表达无影响;（２）ＬＤＬ和ＶＬＤＬ有使这些基因表达增加的趋势;（３）ｏｘ－ＬＤＬ,ｏｘ－ＶＬＤＬ和ｏｘ－ＨＤＬ具有使ＳＭＣｓｉｓ,ｊｕｎ,和ｒａｓ基因表达显著增强的作用（Ｐ＜０．０１）,且其作用较相应的天然脂蛋白大（Ｐ＜０．０１）;（４）天然和氧化修饰型脂蛋白对Ｒｂ基因表达均无影响。据上述结果推测：ＬＤＬ,ＶＬＤＬ,ｏｘ－ＬＤＬ,ｏｘ－ＶＬＤＬ和ｏｘ－ＨＤＬ的致ＡＳ作用可能与刺激ＳＭＣｓｉｓ,ｊｕｎ和ｒａｓ原癌基因表达增加有关。     

鼠伤寒沙门氏菌外膜蛋白诱导BALB／C小鼠免疫保护的实验研究

本实验采用超声破碎、ＴｒｉｔｏｎＸ－１００处理和超速离心技术提取了鼠伤寒沙门氏菌（Ｓａｌｍｏｎｅｌｌａｔｙｈｉｍｕｒｉｕｍ,ＳＴＭ）的外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ,ＯＭＰｓ）,其中脂多糖（ＬＰＳ）的含量约为５％。ＯＭＰｓ经ＳＤＳ—ＰＡＧＥ显示１０余条蛋白带。对ＯＭＰｓ诱发ＢＡＬＢ／Ｃ小鼠产生典型的迟发型变态反应（ＤＴＨ）进行了检测。经腹腔免疫的ＢＡＬＢ／Ｃ小鼠用５００ＬＤ５０鼠伤寒沙门氏菌（５０１１５）攻击,１００％可得到保护;用５００ＬＤ５０伤寒杆菌（Ｅ６８６）攻击     

紫细菌RS601DCPIPH2→MV电子传递的研究

紫色非硫细菌菌株Ｒｂ．ｓｐｈｅｒｏｉｄｅｓ６０１（ＲＳ６０１）的载色体具有典型的Ｒｂ．ｓｐｈａｅｒｏｉｄｅｓ光谱特性。在光照和以ＤＣＰＩＰＨ２为电子供体条件下,ＲＳ６０１载色体能够串联呼吸电子传递链而耗氧或还原ＭＶ而构成ＤＣＰＩＰＨ２→ＭＶ的非循环电子传递链。电子传递抑制剂ＯＰ抑制ＤＣＰＩＰＨ２→ＭＶ电子传递活性,其中Ｉ５０为１．０ｍｍｏｌ／Ｌ;抗霉素Ａ对该电子递影响较小,不存在Ｉ５０。ＬＤＨＸ,     

蟾蜍血清梭曼水解酶（somanase）的纯化及性质的研究

从蟾蜍血清中发现并纯化出了一种与血清载脂蛋白有关的可催化神经性军用毒剂梭曼（０－１,２,２－ｔｒｉｍｅｔｈｙｌｐｒｏｐｙｌｍｅｔｈｙｌｐｈｏｓｐｈｏｆｌｕｏｒｉｄａｔｅ,ｓｏｍａｎ）的水解酶,此酶全部与高密度脂蛋白（ＨＤＬ）以复合形式存在于血清中。我们经过多种纯化手段,从ＨＤＬ中分离纯化出了酯酶的最小分子量组分,得到了高活性、高纯度的酶蛋白,在ＳＤＳ－聚丙烯酰胺凝胶电泳及等电聚焦电泳中呈单一区带,分子量４２ｋＤ,等电点ｐＨ５．２,并证明此酶不是ＨＤＬ中的任何一种已知载脂蛋白,也不是磷脂酶Ａ_２和磷脂酶Ｃ以及存在于ＨＤＬ中的卵磷脂胆固醇酰基转移酶（ＬＣＡＴ）。它是与ＨＤＬ中的主要载脂蛋白ａｐｏＡ_１比较牢固地连接在一起并含量甚微的一种酶蛋白,水解梭曼的Ｋｍ值ｐＨ７．２、３７℃时为３．１３ｍｍｏｌ／Ｌ。可被对氯汞苯甲酸、ＨＣｌ－胍和ＥＤＴＡ－Ｎａ_２所抑制,二硫苏糖醇对酶活性有恢复作用。     

中国大鲵乳酸脱氨酶同工酶研究I组织分布

采用垂直板聚丙烯酰胺凝胶电泳和紫外分光光度法分析大鲵１５种组织的乳酸脱氢酶（ＬＤＨ）同工酶后发现：在正常生理条件下,大鲵体内只有Ｈ４和Ｍ４两种同工酶,红细胞只有Ｍ４一种同工酶;大鲵体内ＬＤＨ同工酶中,以Ｍ４占绝对优势;除生殖腺及脑外,在幼体至成体的发育过程中,ＬＤＨ同工酶未见变化;大鲵体内各组织ＬＤＨ含量及前述两种同工酶相对含量不同。     

瑞氏七鳃鳗乳酸脱氢酶（LDH）同工酶凝胶电泳分析

本文用聚丙烯酰胺凝胶电泳方法,对瑞氏七鳃鳗五种不同组织（骨骼肌、心、肾、肠、鳃）中ＬＤＨ同工酶进行了分析研究,结果表明,ＬＤＨ同工酶具有组织特异性,其中骨骼肌中含有五种ＬＤＨ同。酶,即ＬＤＨ１、ＬＤＨ２、ＬＤＨ３、ＬＤＨ４、ＬＤＨ５,鳃含有ＬＤＨ１和ＬＤＨ４而肾和心只含有ＬＤＨ１,肠只含有ＬＤＨ４。     

中国大鲵乳酸脱氢酶同工酶研究：I组织分布

采用垂直板聚丙烯酰胺凝胶电泳和紫外分光光度法分析大鲵１５种组织的乳酸脱氢酶（ＬＤＨ）同工酶后发现：在正常生理条件下,大鲵体内只有Ｈ４和Ｍ４两种同工酶,红细胞只有Ｍ４一种同工酶;大鲵体内ＬＤＨ同工酶中,以Ｍ４占绝对优势;除生殖腺及脑外,在幼体至成体的发育过程中,ＬＤＨ同工酶未见变化;大鲵体内各组织ＬＤＨ含量及前述两种同工酶相对含量不同。     

沙门菌CWDMs苹果酸脱氢酶的检测

目的：了解沙门菌细菌壁缺陷突变株（CWDMs）的生物氧化及遗传特点和探讨细菌壁缺陷变异的性质与机制。方法：采用PAGE电泳法和分光光度法检测伤寒沙门菌和甲型副伤寒沙门菌及其CWDMs和伤寒沙门菌粗糙型和苹果酸脱氢酶（MDH）同工酶的活性与类型。结果：伤寒沙门菌和甲型副伤寒沙门菌的细菌型和伤寒沙门菌粗糙型经PAGE电泳可见一条MDH同工酶带,CWDMs电泳后可见两条MDH同Ⅰ酶带,在CWDMs的MDH中有一条泳动速率与细菌型及粗糙型的相同,另一条则较快。分光光度法检测证实。细菌型与粗糙型的MDH活性相似,CWDMs的MDH活性则明显较低。结论：CWDMs保留了与亲代细菌型一致的MDH和形成了一种新的MDH,并且其MDH的活性已显著降低,此特性可能与CWDMs生物氧化特性的改变有关。     

A study of enteropathogenic organisms isolated in the public health laboratory, Lusaka.

A total of 328 specimens of stools were examined in the Public Health Laboratory during January and February 1973. Enteropathogens were isolated from 117 of these specimens. Besides these, 12 strains of Salmonellae were isolated from blood and 8 from urine. An occasional Salmonella was isolated from the pleural fluid (S. paratyphi A) pus from the knee (S. enteritidis) and from the C.S.F. of an infant (S. paratyphi C.). Salmonella typhi and Salmonella paratyphi A are the predominant Salmonella species. No Salmonella paratyphi B has been isolated. Shigella, was isolated with slightly less frequency than Salmonella, and Shigella flexneriis was the predominant species. E. coli 0112/K66 is the most common enteropathogenic E. coli. The majority of the Shigella and Salmonella species are sensitive to the common antibiotics used. The E. coli organisms show multiple resistance to a number of antibiotics.     

Genetics of Sensitivity of Salmonella Species to Colicin M and Bacteriophages T5, T1, and ES18

Nearly all of 62 strains of Salmonella paratyphi B were sensitive to colicin M and phage T5 but resistant to phages T1 and ES18 and to colicin B. All tested S. typhimurium strains were resistant to colicin M and phage T5, and many were sensitive to phage ES18. A rough S. typhimurium LT2 strain given the tonA region of Escherichia coli or S. paratyphi B became sensitive to colicin M and phage T5. We infer that the tonA allele of S. paratyphi B, like that of E. coli, determines an outer membrane protein that adsorbs T5 and colicin M but not phage ES18, whereas the S. typhimurium allele determines a protein able to adsorb only ES18. The partial T1 sensitivity of a rough LT2 strain with a tonA allele from E. coli or S. paratyphi B and also the tonB(+) phentotype of an E. coli B trp-tonB Delta mutant carrying an F' trp of LT2 origin showed that S. typhimurium LT2 has a tonB allele like that of E. coli with respect to determination of sensitivity to colicins and phage T1. Rough S. paratyphi B, although T5 sensitive, remained resistant to T1 even when given F' tonB(+) of E. coli origin. Classes of Salmonella mutants selected as resistant to colicin M included: T5-resistant mutants, probably tonA(-); mutants unchanged except for M resistance, perhaps tolerant; and Exb(+) mutants, producing a colicin inhibitor (presumably enterochelin). Some Exb(+) mutants were resistant to a bacteriocin inactive on E. coli but active on all tested S. paratyphi B and S. typhimurium strains (and on nearly all other tested Salmonella). A survey showed sensitivity to colicin M in several other species of Salmonella.     

Antigen sharing of Salmonella typhi non-flagellar proteins with other salmonellae and some shigellae and Escherichia coli

Cathodal moving protein components were identified in agarose gel electrophoresis of the Veronal buffer extract of a non-motile strain of S. typhi (8393, Colindale). Rabbit antiserum was raised against these cationic proteins; it had both agglutinating and precipitating activity. A total of 80 salmonella strains belonging to serogroups A, B, C1, C2, D, E1 and E2 including 26 S. typhi and 10 S. paratyphi A were tested against this antiserum in a slide agglutination test; all strains were agglutinated. Among 94 other bacterial strains tested, the antiserum agglutinated all 16 strains of Shigella flexneri, 2 of 5 Shigella sonnei, 5 of 34 E. coli and 1 of 8 Citrobacter species. These results show that there is antigenic sharing of the non-flagellar proteins of S. typhi with many other salmonellae as well as with some shigellae and E. coli.     

Delayed hypersensitivity in murine salmonellosis: specificity of footpad reaction in mice infected with rough mutants of Salmonella typhimurium

Delayed type (footpad) hypersensitivity (DTH) in BALB/c mice immunized with rough mutant strains of Salmonella typhimurium LT2 was examined. Injection of live organisms of an Rb mutant TV148 strain induced DTH in mice, while injection of the heat-killed organisms did not. The mice immunized with live organisms of the Ra, Rb, Rc, Rd, and Re mutant strains showed positive footpad reactions to the heat-killed cell antigen of LT2 (wild type) strain. The mice immunized with the Rb mutant strain also showed positive footpad swellings in response to heat-killed cell antigens of S. paratyphi A, S. paratyphi B, S. typhi, S. enteritidis, and S. cholerae-suis. Furthermore, positive reactions to antigens of Escherichia coli and Shigella flexneri were seen in the TV148-immunized mice, but the mice did not respond to heat-killed organisms of Pseudomonas aeruginosa or Staphylococcus aureus. The cross-reactive footpad reaction to E. coli could be transferred adoptively with T cells prepared from the spleens of TV148-immunized mice into syngeneic recipients. These results suggest that the cross-reactive DTH antigen(s) is widely distributed among related organisms such as Shigella and Escherichia.     

Surveillance for typhoid fever in Matsuyama city during 1974-1981 and detection of Salmonella typhi in sewage and river waters

Eighty-two cases of typhoid fever were found in Matsuyama city in the period from 1974 to 1981. Seventy-six cases were found to be infected with Salmonella typhi other three with Salmonella paratyphi A, and the remaining three were diagnosed only clinically. The strains of S. typhi isolated from these patients showed such a variety of Vi-phage types as D1, D2, E1, M1, 53 and degraded Vi-positive strain (DVS). The concurrent survey of the city sewage and river waters for typhoid bacilli was conducted with total 578 samples taken therefrom. S. typhi was isolated from 120 of those samples. The Vi-phage types of the isolates were closely related with those of the isolates from the patients. The periodical examinations of the city sewage and the draining river may serve as a useful means for the controlling typhoid fever epidemics.     

Bafoudiosbulbins A, and B, two anti-salmonellal clerodane diterpenoids from Dioscorea bulbifera L. var sativa

Two clerodane diterpenoids, Bafoudiosbulbins A 1, and B 2, together with five known compounds: tetracosanoic acid, 1-(tetracosanoyl)-glycerol, trans-tetracosanylferulate, beta-sitosterol and 3-O-beta-D-glucopyranosyl-beta-sitosterol were isolated from the tubers of Dioscorea bulbifera L. var sativa. Their structures were established by spectroscopic methods (1D and 2D-NMR, MS) and X-ray crystallographic diffraction analysis of compound 1. The CH2Cl2-soluble portion of the crude extract and the two clerodanes were screened for anti-bacterial activity using both agar diffusion and broth dilution techniques against Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella typhi, Salmonella paratyphi A and Salmonella paratyphi B. They both showed significant activities against P. aeruginosa, S. typhi, S. paratyphi A and S. paratyphi B.     

6种沙门菌单克隆抗体的制备和效能评价

目的：制备稳定、特异、高亲和性的分别针对甲型副伤寒沙门菌、乙型副伤寒沙门菌、丙型副伤寒沙门菌、肠炎沙门菌、伤寒沙门菌和猪霍乱沙门菌的单克隆抗体。方法：用甲醛灭活的菌液抗原免疫BALB／c小鼠,取脾细胞与SP2／0骨髓瘤细胞融合;用灭活的菌液包被酶标板,ELISA筛选阳性克隆株,建立细胞系;选取高效分泌杂交瘤细胞,常规制备腹水并纯化,进行单抗特异性与亲和性评价。结果：筛选得到分泌6种沙门菌相应单克隆抗体的杂交瘤细胞株,获得高亲和性单抗;所有单抗与大部分病原菌（包括7种沙门菌、3株志贺菌、2株李斯特菌、4株致病性大肠杆菌、2株霍乱弧菌）无交叉反应,但由于同类型O抗原的广泛分布,抗乙型副伤寒沙门菌单抗与鼠伤寒沙门菌、抗伤寒沙门菌单抗与肠炎沙门菌有明显的交叉反应。结论：沙门菌单抗的制备,为感染性腹泻的监测、诊断奠定了基础。     

Genetic Basis of Vi Antigen Expression in Salmonella paratyphi C

Analysis of hybrids formed in a cross between a Salmonella paratyphi C Hfr and an S. typhimurium recipient indicated that the structural genetic determinants of the S. paratyphi C Vi antigen are located closely adjacent to the mel determinant, between this marker and purA. A similar location was indicated for the structural genetic determinants of the S. typhi Vi antigen (the viaB locus) by the results of a mating in which a hybrid S. typhimurium Hfr bearing the S. typhi viaB determinants was used to transfer these genes to an S. typhimurium recipient. Mating experiments with a Vi-antigen-expressing S. typhi Hfr and an S. typhimurium hybrid recipient expressing the Vi antigen of S. paratyphi C yielded no recombinants in which loss of Vi antigen expression occurred, indicating that the chromosomal locus occupied by the genetic determinants of the S. paratyphi C Vi antigen is the same one at which, in S. typhi, the viaB genes reside. Introduction of a mutant S. typhi viaA gene into an S. typhimurium hybrid expressing the Vi antigen, as the consequence of prior receipt of the S. paratyphi C viaB determinants, resulted in that hybrid's loss of Vi antigen expression, demonstrating that the viaA determinant plays a role in Vi antigen expression in S. paratyphi C, as well as in S. typhi. Although the percentages of coinheritance of the viaB and mel determinants in the mating experiments suggested that their linkage is sufficiently close to allow cotransduction by P22, attempts to accomplish this with lysates prepared on S. typhimurium hybrids expressing either S. typhi or S. paratyphi C viaB determinants were not successful.