慢性肾盂肾炎L型细菌感染及耐药分析

目的了解L型细菌在慢性肾盂肾炎的感染及耐药状况。方法对71例患者清洁中段尿做普通细菌培养(B型)、L型细菌培养(L型)及耐药分析。结果细菌阳性率为77.5%,其中单独L型阳性率为49.3%、B型与L型混合感染为15.5%,而B型阳性率仅为12.7%。主要是大肠埃希菌,其次是葡萄球菌;青霉素及头孢噻肟均有较高的耐药率(88.9%及73.6%)。结论L型细菌在慢性肾盂肾炎感染中占主导,β-内酰胺类药物有较高的耐药性,临床治疗应据药敏结果合理选择及时调整抗生素。     

咽部活组织中细菌L型的检出及意义

应用病原微生物培养、电镜、组织切片细菌学检查及Ｌ型抗体免疫组化染色等方法,检测６４例慢性咽炎组织的细菌Ｌ型。结果有４２例培养出细菌Ｌ型,其阳性率为６５．６％;它与切片革兰氏染色Ｌ型检出阳性率（６７．２％）无显著性差异,Ｐ＞０．０５。透射电镜在慢性咽炎组织的间质及上皮细胞、巨噬细胞等细胞内见皮细菌Ｌ型;且Ｌ型抗体免疫组化染色亦证实组织中有细菌Ｌ型抗原。提示,细菌Ｌ型感染与慢性咽炎关系密切,Ｌ型侵入组织并在宿主细胞内生长的特征,可能是慢性咽炎反复发作,迁延不愈的重要原因。     

Ultrastructure of L forms. IV. L forms of nonagglutinating vibrios]

A study was made of the ultrastructure of stable L-forms of Nag vibrios aged 24 hours. Cells of all types of the L-forms had cytoplasmic membranes, and a three-layered structure, which was found not everywhere. Externally of the cytoplasmic membrane, in some areas of the individual cells there were revealed a plastic layer of cell wall and a basal membrane. However, in difference to bacterial forms of the vibryos, rigidity of the cell wall was disturbed, and the links between the cell wall and the cytoplasmic membrane were indetectable. There were regularly revealed lamellar of myelin-like membranous structures in the cytoplasm, which did not occur in bacterial forms, and also lamellar mesosomes. The latter were found in the sites of cell division. Viability of small bodies as the minimal reproductive forms of the L-cultures is confirmed by the presence in them of a nucleoid and of the binary division.     

Isolation of L-Forms in a Clinical Microbiology Laboratory

Previous studies have demonstrated that L-forms of bacteria may play a role in persistent, chronic, or recurrent urinary-tract infections. A 2-year program was initiated to determine the feasibility of culturing for L-forms on a routine basis, and to determine the effectiveness of such a program. In relation to the total number of specimens, few L-forms were actually isolated. In comparison with the amount of equipment and technician time required, the return was negligible; only 0.5% of all urine specimens were positive for L-forms. An increase to only 1.2% was noted when culturing for L-forms was limited to patients with a diagnosis of bacteriuria or pyelonephritis. It is recommended that this technique be reserved for those patients with a long history of recurrent urinary-tract infections, after other attempts to cure the patient have met with failure.     

Morphologic characteristics of spheroid elements in a culture of bacterial L-forms according to scanning electron microscopy findings]

The authors studied stable L-cultures of Proteus valgaris, Bac. subtillis, Staphylococcus aureus, Streptococcus pyogenes of group A, and also unstable cultures of the L-forms of Proteus vulgaris and Proteus vulgaris culture at the stage of spheroplasts. Spheroid cells proved to appear at the stage of spheroplasts, prevailed at the log phase in stable and unstable L-cultures, but were less frequent at the stationary phase. Cross section of L-colonies showed that they were located at the surface. The size of spheroid elements was from 1 to 5 micron; their surface was smooth or slightly wrinkled with numerous protrusions and individual sockets. The spheroid cells were distributed in the colonies freely, in clusters, or were connected to one another by anastomosis. Several methods of reproduction of spheroid cells are described, including equal and unequal binary fission, budding, and formation of elementary bodies within the cell. Morphological connection of spheroid cells with large bodies, filamentous structures and structureless matrix of the L-colony apparently pointed to their origin from the corresponding elements of the L-cultures.     

人体病理组织中细菌L型的电镜观察

对9例细菌L型感染的人体病理组织进行透射电镜观察。结果显示:(1)L型可分布于组织间质或进入吞噬细胞、上皮细胞和癌细胞等细胞胞质;(2)L型具有多形态、大小不等、胞壁缺陷、电子密度低等特点,细胞胞质内的细菌L型尚有A、B两种形态区别;(3)L型在组织中形成了不完全箍缩分裂;(4)宿主细胞超微结构仅出现轻微改变。本结果与文献报道基本一致,提出了透射电镜下病理组织中细菌L型的形态特征及分布;并对L型感染与慢性炎症的关系等问题进行了讨论。     

Microbial associations in chronic respiratory tract diseases

In the peripheral blood of patients with chronic bronchitis the presence of Mycobacterium pneumoniae antigens has been registered in 20.0% of cases and the presence of group A Streptococcus haemolyticus antigens, in 24.0% of cases, the transformation of streptococci into the L-form being observed in 12.0% of cases. The presence of M. pneumoniae, streptococci and their L-forms, as well as associations of these microorganisms, is characteristic of patients with chronic obstructive bronchitis, which is, probably, one of the reasons for the maintenance of bronchial obstruction. In patients with chronic pneumonia and fibrosing alveolitis the antigens of the bacterial form of streptococcus and its L-forms have been detected only in a few cases.     

L-Form Induction, Morphology, and Development in Two Related Strains of Erysipelothrix rhusiopathiae

Two related strains of Erysipelothrix rhusiopathiae, one the parent and the other an L-form revertant, were studied for their propensity or ability to produce L-forms under the influence of penicillin. The parent strain produced L-forms in nutrient solid media in an osmolarity range between 0.85 and 5.0% NaCl concentration whereas the revertant strain did so between 0.5 and 3.0% NaCl concentration. When various hyperosmolar media were tried without penicillin, recovery of L-forms from the revertant strain was optimal at a salt concentration of 2.0%, whereas the parent strain occasionally produced a few L-forms on 3.0% salt medium only. The process of penicillin-induced transformation from bacteria to L-form followed an unusual morphological sequence, beginning with beading of the bacterial body, followed by disintegration into granules from which the L-form colony derived. No large bodies were seen during the initial process of L-form induction, but they evolved later from the original granules and had the potential to reproduce L-type growth. The spontaneous development of L-forms in hyperosmolar media had a different morphological sequence starting with elongation of the bacteria into filaments which later developed polar and central dilatations from which granules and L-type growth developed. The differences in biological behavior between these related bacterial strains suggest that the revertant strain developed new properties, probably of genetic origin. Consequently, the assumption that L-forms revert to the "parent" bacteria may not always be justified. It can be made only after the biological properties of the parent and the revertant organisms have been properly identified.     

The role of Tamm-Horsfall protein in the pathogenesis of reflux nephropathy and chronic pyelonephritis

Recurrent bacterial infection of the kidney was previously thought to be responsible for the renal scarring typical of chronic pyelonephritis until recent studies suggested that recurrent bacteriuria rarely produces chronic pyelonephritis in the absence of obstructive uropathy. In contrast, the association between vesicoureteral reflux (VUR) and chronic pyelonephritis has been observed frequently in the absence of urinary infection. Although the mechanism by which VUR injures the kidney has not been defined, recent observations have suggested that some component of urine might serve as an antigenic determinant involved in the immunopathogenesis of renal scarring in VUR. Therefore, the present studies investigated the immunopathogenic role of Tamm-Horsfall protein (THP) in (1) a rabbit model of tubulointerstitial nephritis; (2) a swine model of reflux nephropathy; and (3) patients with recurrent nephrolithiasis. The antigenic similarities between THP and uropathic bacteria were also studied. Our observations indicate that autoimmune responses to THP may occur after exposure to THP by intravenous challenge in rabbits, by urinary reflux in pigs, and in recurrent nephrolithiasis in man. Also, extracts of uropathic coliforms competitively inhibit the binding of human THP to its antibody. These studies suggest that autoimmune responses to THP may be the pathogenetic mechanism by which these factors, including bacteriuria, contribute to "chronic pyelonephritis."     

四种细菌L型的形态定量分析

用抗生素或臭氧的方法诱导出四种细菌和Ｌ型后,用图像分析技术定量检测Ｌ型多形性,表面积、最大直径、周长、等效圆直径和形状因子,并作比较分析。四种细菌Ｌ型及Ｌ型中的巨形体在上述５项形态学参数指标上除形状因子外均较细菌型明显增大,而形状因子参数明显小于细菌型。细菌Ｌ型之间在形态学指标上的差异有显著性（ｐ＜０．０５）。形态定量分析能准确客观地反映形态定性观察的结果以及细菌Ｌ型的形态变异程度,为细菌Ｌ型的形态定量分析提供依据。     

Serum and urine levels of interleukin-6 and interleukin-8 in children with acute pyelonephritis

Urinary tract infection (UTI) is a common clinical disorder in younger infants and children and may result in permanent renal damage. The inflammatory cytokines interleukin (IL)-6 and IL-8 play an important role in response to bacterial infection. This prospective study investigated the association between serum and urine IL-6 and IL-8 levels and acute pyelonephritis confirmed by (99m)Tc-dimercaptosuccinic acid (DMSA) scan. A total of 78 children aged 1-121 months with a diagnosis of first-time febrile UTI were included. The following inflammatory markers were assessed: fever; white blood cells count (WBC); C-reactive protein (CRP); and serum and urine IL-6 and IL-8. The patients were divided into the acute pyelonephritis group (n=42) and the lower UTI group (n=36) according to the results of DMSA scan. Fever, WBC and CRP levels were significantly higher in children with acute pyelonephritis than in those with lower UTI (all p <0.001). Significantly, higher initial serum and urine IL-6 and IL-8 levels were found in children with acute pyelonephritis than in those with lower UTI (all p <0.001). Serum and urine IL-6 in children with acute pyelonephritis were positively correlated with fever, CRP and leucocyturia. These results indicate that both serum and urine IL-6 and IL-8 levels, particularly IL-6, are useful diagnostic tools for early recognition of acute pyelonephritis in febrile children.     

金黄色萄葡球菌L型特异性抗体的免疫组化试验在妇科感染性疾病诊断上的应用

本文对49例慢性子宫内膜炎、慢性宫颈炎等妇科疾病的病理切片,用金黄色葡萄球菌细菌型及L型的特异性杭体作酶免疫组化染色(PAP法)证明在金黄色葡萄球菌引起的各种妇科感染中,细菌型及L型混合感染占60％,单独细菌型感染占28.9％,单独L型感染占11.1％。本研究证明了金葡菌变异为L型后,有特异性杭原出现,并证明细菌L型的特异性抗原有一定的免疫学诊断价值。     

鼻咽癌与细菌L型的关系

大量研究表明,鼻咽癌与ＥＢ病毒有关。通过对９８例鼻咽癌组织的切片革兰氏染色Ｌ型检查、电镜和Ｌ型抗体免疫组化染色等研究,发现鼻咽癌组织中细菌Ｌ型亦甚常见,切片革兰氏染色有７８例查见细菌Ｌ型,其阳性率为７９．６％;免疫细化染色Ｌ型抗原检出阳性率为６２．２％,电镜不仅在细胞间质见到细菌Ｌ型,而且在癌细胞、巨噬胞细等细胞胞质内也见到细菌Ｌ型。提示,细菌Ｌ型与鼻咽癌关系十分密切,很可能是鼻咽癌致癌因子之一。     

3种革兰氏阴性细菌及其L型内毒素含量的测定

本文采用鲎试剂对大肠杆菌（ＡＴＣＣ２５９２２）、伤寒杆菌、绿脓杆菌（ＡＴＣＣ７８５３）及它们的Ｌ型内毒素的含量进行测定。结果显示细菌型与细菌Ｌ型均具有内毒素,但细菌Ｌ型内毒素含量较细菌型低（约为１／３￣１／２）。因此,认为细菌Ｌ型仍有一定的致病性。     

细菌L型与喉癌关系的研究

用组织切片革兰氏染色、免疫组织化学染色等方法,对８５例喉癌重新切片,进行细菌Ｌ型检查,结果发现有６５例革兰氏染色Ｌ型菌阳性,其阳性率为７６．５％。５３例（６４．７％）Ｌ型抗体免疫组化染色和革兰氏染色Ｌ型菌均阳性,两者总符合率为８３．５％。细菌Ｌ型呈多形性,分布于癌巢、癌间质,常聚集成堆,也可呈散在性分布。５８例（６８．２％）癌细胞胞浆内也见到Ｌ型菌。提示细菌Ｌ型感染与喉癌关系密切。     

Bacterial L-forms require peptidoglycan synthesis for cell division

Cell-wall-less bacterial variants, or L-forms, have been described in many bacterial species under laboratory conditions, in infected eukaryotic cell cultures and inside animals. Of special interest for human health is the formation of L-forms as a consequence of specific antibiotic treatments, and the potential involvement of L-forms in persistent and relapsing infections. An old enigma about L-forms is how they can divide in the absence of cell wall synthesis, since septum formation is an essential requisite for cell division. However, the classical definition of L-forms as cell-wall-less bacterial variants may need a revision to accommodate recent observations by Richard d'Ari and coworkers: genetic and biochemical evidence indicates that E. coli L-forms induced by beta-lactam antibiotics do contain small amounts of peptidoglycan, essential for their growth and probably required for septum formation. If these observations are extrapolated to all known L-forms, the very concept of cell-wall-less bacteria may need revision, and be restricted to mycoplasmas and their relatives.     

Processes of bacterial L transformation and L form reversion in the body of argasid ticks

The experimental infection of tampan ticks (Ornithodoros moubata) with the bacterial cultures of Listeria monocytogenes and Salmonella typhimurium, as well as with their L-forms, was carried out. These experiments demonstrated that both the L-transformation of bacteria and the reversion of their L-forms into the initial bacterial culture could occur in the body of the ticks.     

血平板针尖样及云雾状菌落分离与鉴定

采集98份临床标本接种血平板培养,40份生长出针尖样及云雾状菌落。经分离鉴定38株为细菌L型,其中金黄色葡萄球菌（Staphyloeoccusaureus）居首位,占52．1％（20／38）．其返视性与L型平板分离相比较差。但对抗生素的敏感性则无差别。表明血平板可用于分离细菌L型,并保持了传统L型的特性。     

ExperimentalEscherichia coli pyelonephritis: Formation of spheroplasts in the kidney of the untreated rat

An experimental hematogenous pyelonephritis produced in the rat byEscherichia coli was studied from the point of view of the possible rôle of L-forms of bacteria in chronic renal infection. Osmotically fragile cells were shown in the kidneys from the 3rd day after inoculation, and their percentage of the totalE. coli cell number increased during the course of the infection. In order to distinguish bacteria and spheroplasts of theE. coli strain used, two more methods were elaborated: 1) a bacteriological method based on the different sensitivity of bacteria and spheroplasts to erythromycin; this method confirmed the results obtained with osmotic shock; 2) an immunological method, using the indirect fluorescent antibody technique and an antiserum specifically labelling the spheroplasts; in kidneys removed 10–15 days after the inoculation, spherical bodies with morphological and antigenic characteristics similar to those of spheroplasts became visible.     

正常人和肝病患者血中“致密体”及其与纳米细菌的关系

目的研究正常和肝病患者血液中圆球体样微生物的生物学特性,初步确定纳米细菌与该种可滤过物的关系。方法将正常人和肝病患者培养阴性的血培养物进行细菌L型、穿菌和厌氧菌培养。同时取沉淀用透射电镜观察。L型培养液用0．45μm和0．22μm滤器过滤,接种RPMI 1640培养基,细胞培养条件下培养45d,用鼠抗纳米细菌单克隆抗体8D10免疫组化和钙盐染色进行纳米细菌鉴定。血浆纳米细菌培养阳性者12000×g离心后,进行普通和L型细菌培养。结果36／39患者和60／60健康对照血培养液中呈现类似于L型的巨型体、圆球体、原生小体的不明微生物。电镜观察圆球体内为电子致密样物质,周围未见细胞壁结构。电镜和光镜下可见其粘附在红细胞上或存在于红细胞内。滤过后培养物钙盐染色有5／39阳性,但纳米细菌免疫组化染色均阴性。纳米细菌阳性的培养物转种,未见一般细菌、细菌L型和上述血中“致密体”。结论血培养中致密体样微生物与纳米细菌无关,可能在维持机体正常免疫功能上具有一定意义。