幽门螺杆菌内毒素对小鼠成纤维细胞和人胃粘膜组织细胞的侵入和侵害

应用免疫胶金技术证实幽门螺杆菌内毒素作用于小鼠成纤维细胞后５ｍｉｎ即能侵入胞浆，６０ｍｉｎ后可在细胞核中检出ＨＰ－ＬＰＳ结合的胶合颗粒，免疫组织化学法检测结果表明，６５．４％，胃粘膜活检标本组织吸ＨＰ－ＬＰＳ存在，受ＨＰ－ＬＰＳ作用的小鼠成纤维细胞和ＨＰ－ＬＰＳ检测阳性的胃炎患者活检标本细胞均出现相似的超微结构病变，如粗面内质网扩张，膜旁核糖体颗粒脱落，线粒体嵴或外膜消失等。实验结果表明幽门螺杆菌     

钩端螺旋体脂多糖对小鼠成纤维细胞和豚鼠脏器组织的侵入及损害

应用免疫胶金技术证实钩端螺旋体脂多糖（Ｌ－ＬＰＳ）作用于体外培养的小鼠成纤维细胞后５ｍｉｎ即能侵入胞浆,３０ｍｉｎ后可在细胞核中检出Ｌ．一！Ｉ５。钩端螺旋体实验感染的豚鼠各内脏组织的细胞间隙、胞浆及核等内均存在１．一！,ＰＳ,尤以肾和肺脏检出阳性率较高。Ｌ－ＬＰＳ检测阳性的小鼠成纤维细胞和感染豚鼠内脏组织细胞均出现相似的超微结构病变,如内质网扩张、膜旁核糖体脱落、线粒体嵴或外膜消失等。实验结果表明Ｌ－ＬＰＳ是钩端螺旋体主要致病物质之一。     

钩端螺旋体脂多糖对小鼠成纤维细胞和豚鼠脏器组织的侵入及…

应用免疫胶金技术证实钩端螺旋体砩多糖作用于体外培养的小鼠成纤维细胞后５ｍｉｎ即能侵入胞浆，３０ｍｉｎ后可在细胞核中检出Ｌ－ＬＰＳ。钩端螺旋体实验感染的豚鼠各内脏组织的细胞间隙，胞浆及核等内均存在Ｌ－ＬＰＳ，尤以肾和肺脏检出阳性率较高。     

幽门螺杆菌脂多糖的细胞毒性及其抗体保护作用的研究

幽门螺杆菌脂多糖的细胞毒性及其抗体保护作用的研究黄醴森，方平楚，严杰近年来，一些学者的研究结果证实幽门螺杆菌－脂多醣（Ｈｐ－ＬＰＳ）具有细菌内毒素的生物学活性［１～３］。我们应用免疫组化和３Ｈ－ＴｄＲ掺入等方法，研究了慢性胃炎胃粘膜标本中Ｈｐ－ＬＰＳ...     

幽门螺杆菌内毒素的化学组成和结构的初步分析

幽门螺杆菌内毒素的主要化学成份为糖类和脂类,其戊糖、己糖、庚糖和氨基糖的含量分别为13.2％、25.6％、9.3％和5.2％(w/w)。幽门螺杆菌内毒素中所含的2—酮—3—脱氧—D—甘露糖—辛酮糖酸(KDO)含量高达7.2％(W/W)。该内毒素的红外光谱和超微结构与大肠杆菌内毒素十分相似。电镜下,幽门螺杆菌内毒素呈细长的条索状结构。实验结果表明,幽门螺杆菌内毒素的化学组成和超微结构与大肠杆菌内毒素基本相同。     

酶联免疫法检测粪便幽门螺杆菌特异性抗原

目的：探讨酶联免疫法（ＥＬＩＳＡ）检测粪便幽门螺杆菌（ＨＰ）特异性抗原（ＨＰＳＡ）在诊断ＨＰ感染中的价值。方法：９９例患者通过快速尿素酶试验及组织学Ｗａｒｔｈｉｎ－Ｓｔａｒｒｙ银染法检测，分为ＨＰ感染组和非感染组，用酶联免疫法检测粪便中ＨＰＳＡ。结果：粪便酶联免疫法检测ＨＰ感染的敏感性为１００％，特异性为８５．７１％。结论：酶联免疫法检测粪便中ＨＰＳＡ敏感性、特异性高，且取材方便，方法简单，适合临床应用。     

幽门螺杆菌感染BALB/c无菌小鼠的免疫应答

目的 探讨幽门螺杆菌感染步鼠的免疫就应答状况。方法 建立ＨＰ经口感染无菌小鼠的动物模型,设立感染免疫组、感染对照组与免疫对照组３组,分别进行试验。结果 感染免疫组抗ＨＰ的特异性ＩｇＧ有显著增高,且该组小鼠胃中的ＨＰ菌数大量减少。结论此种免疫应答以体液免疫为主,细胞免疫所占比例很少,不能彻底清除小鼠胃中的ＨＰ。     

聚合酶链反应对胃活检标本中幽门螺杆菌DNA的检测

应用聚合酶链反应（ＰＣＲ）对幽门螺杆菌（ＨＰ）特异性的尿素酶Ａ基因进行扩增，４０个循环可以检测出１０个ＨＰ，加用非同位素地高辛探针杂交，可以检出单个ＨＰ。用本法对１００例胃镜活检标本进行ＰＣＲ检测，其阳性率达８２％，而尿素酶水解试验，细菌涂片及培养的阳性率则分别为６８％、４６％、１８％。ＰＣＲ方法能快速、敏感，特异地检测出临床标本中ＨＰ，对于研究ＨＰ与上消化道疾病之间的关系，指导抗菌治疗均具有重要意义。     

聚合敏链反应对胃活检标本中幽门螺杆菌DNA的检测

应用聚合酶链反应（ＰＣＲ）对幽门螺杆菌（ＨＰ）特异性的尿素酶Ａ基因进行扩增，４０个循环可以检测出１０个ＨＰ，加用非同位素地高辛探针杂交，可以检出单个ＨＰ。用本法对１００例胃镜活检标本进行ＰＣＲ检测，其阳性率达８２％，而尿素酶水解试验，细菌涂片及培养的阳性率则分别为６８％、４６％、１８％。ＰＣＲ方法能快速、敏感、特异地检测出临床标本中ＨＰ，对于研究ＨＰ与上消化道疾病之间的关系，指导抗菌治疗均具有重要     

诱生型一氧化氮合成酶在慢性胃炎组织中的表达及其与HP感染的关系

为了探讨诱生型一氧化氮合成酶（ｉＮＯＳ）在慢性胃炎组织中的表达强度及其与幽门螺杆菌（ＨＰ）感染的关系，我们应用免疫组化法对６例正常对照者及５６例慢性胃炎患者胃组织病理标本的ｉＮＯＳ表达强度进行检测，并用改良Ｇｉｅｍｓａ法同步检测其ＨＰ感染状况。现报告...     

Detection of H.pylori DNA in gastric epithelial cells by in situ hybridization

AIM: To investigate the presence of H.pylori DNA within gastric epithelial cells in patients with H.pylori infection and its possible carcinogenic mechanism. METHODS: Total 112 patients, with pathologically confirmed chronic superficial gastritis, chronic atrophic gastritis, intestinal metaplasia, atypical hyperplasia or gastric cancer were studied. Among them, 28 were H.pylori negative and 84 H.pylori positive. H.pylori DNA in gastric epithelial cells was detected by GenPoint catalyzed signal amplification system for in situ hybridization. RESULTS: In the H.pylori positive group, zero out of 24 chronic superficial gastritis (0.0%), four out of 25 precancerous changes (16.0%) and thirteen out of 35 gastric cancers (37.1%) showed H.pylori DNA in the nucleus of gastric epithelial cells, the positive rates of H.pylori DNA in the nucleus of gastric epithelial cells were progressively increased in chronic superficial gastritis, precancerous changes and gastric cancer groups (chi(2)=12.56, P=0.002); One out of 24 chronic superficial gastritis (4.2%), eleven out of 25 precancerous changes (44.0%) and thirteen out of 35 gastric cancers (37.1%) showed H.pylori DNA in the cytoplasm of gastric epithelial cells (chi(2)=10.86, P=0.004). In the H.pylori negative group, only one patient with gastric cancer was found H.pylori DNA in the nucleus of gastric epithelial cells; Only two patients, one patient with precancerous changes and another with gastric cancer, showed H.pylori DNA in the cytoplasm of gastric epithelial cells. Furthermore, H.pylori DNA must have been in the cytoplasm as long as it existed in the nucleus of gastric epithelial cells. CONCLUSION: H.pylori DNA exists both in the nucleus and the cytoplasm of gastric epithelial cells in patients with H.pylori infections. The pathological progression from chronic superficial gastritis, precancerous changes to gastric cancer is associated with higher positive rates of H.pylori DNA presence in the nucleus of gastric epithelial cells.     

Detection of cytoplasmic and surface membrane markers in cells of some human hematopoietic cell lines.

The cells of some human leukemia-lymphoma T cell lines (JURKAT, MOLT4), B cell lines (DAUDI, U-266) and of myeloid U-937 cell line were characterized for their surface membrane and cytoplasmic marker profiles. The usefulness of some fixation and permeabilization methods of cell membrane for detection of cytoplasmic markers by flow cytometry was studied. The methods of cell fixation in suspension were found to be more sensitive than the methods of cell fixation in smears. With the very short buffered formaldehyde-acetone (BFA) fixation used in this study an optimal penetration of the monoclonal antibodies (MoAbs) through the plasma membrane and specific binding to the appropriate structures were achieved. CD22 antigen was detected in cytoplasm but not on membrane of DAUDI cells. In another B cell line, U-266, CD22 antigen was present both in cell membrane and cytoplasm. The marker corresponding to anti-CD19 MoAb was detected in cytoplasm but was absent on membrane of U-266 cells. Furthermore, the antigen estimated by anti-CD3 MoAb could be detected intracellularly in cells of both T cell lines tested, while it was absent on cell membrane of these cells. The phenotypic study of U-937 cells showed that the majority of cells expressed myeloid associated antigens. In our study the CD14 marker detected on cell surface membrane of U-937 cells was missing in their cytoplasm. The surface antigens remained intact after BFA fixation enabling a simultaneous detection of membrane and cytoplasmic markers in double immunofluorescence studies. Through this combination of markers minor cell populations could be detected. Human hematopoietic cell lines could serve as a reliable model system for a rapid and quantitative immunodiagnosis.     

Ultrastructure of endocrine-like cells in lamina propria of human gastric mucosa.

Endocrine cells of gastric and gut mucosa are commonly thought to be present only within mucosal glands. In a previous report, we described argyrophilic cells in the lamina propria in 40% of surgical gastric specimens, using light microscopy. All these patients had chronic gastritis. Argyrophilia, however, is a non-specific reaction which could occur in other than endocrine cells. The present study was undertaken to describe the ultrastructure of argyrophil cells in the lamina propria. In five patients with chronic gastritis, endoscopic biopsies were taken from the fundic, intermediate, and pyloric areas of the stomach. Single and/or clustered argyophil cells were seen by light microscopy in the lamina propria of the intermediate and pyloric areas. On electron-microscopy, these cells had the following characteristics of endocrine-like cells: they were characterised by numerous electron dense granules in the cytoplasm, 100-300 nm in diameter; the cytoplasm contained poorly-developed rough endoplasmic reticulum and well-developed smooth endoplasmic reticulum with occasional vesicles. Immunostaining gave negative results for various gastrointestinal hormones. These ultrastructural characteristics of lamina propria cells are similar to endocrine cells of the APUD series. We conclude that endocrine-like cells occur in the lamina propria of the human stomach in the presence of chronic gastritis.     

Ultrastructural observation of Helicobacter pylori to the gastric epithelia in chronic gastritis and peptic ulcers

AIM: The relationship between Helicobacter pylori (Hp) and gastric epithelia in chronic gastritis and in peptic ulcers was studied by transmission electron microscopy (TEM). METHODS: Seventy-five patients were screened for Hp. Gastric antral biopsy specimens were fixed in glutaraldehyde and treated with tannic acid before OsO₄ staining. Samples were routinely processed for TEM studies (at least four semi-thin sections oriented for ultrathin sections in each sample). RESULTS: The bacilli were detected by TEM within the gastric mucosa in 53 of 55 patients infected with Hp. Ultrathin sections revealed clear glycocalyx by which the bacillus was connected to the epithelium. As the bacilli colonized, the adjacent mucous cells degenerated. They were characterized by erosion of the juxtaluminal cytoplasm, vacuolation or blebbing, and desquamation of the cell membrane. The bacilli located in the lumen attracted neutrophils, which migrated into intercellular space of the epithelia or into the lumen to begin phagocytosis of Hp. CONCLUSION: The sensitivity and specificity of TEM diagnosis is 96% and 95%, respectively. Tannic acid is suitable for the preservation of the glycocalyx of a cell. The colonized bacilli, usually with the wide periplasm, contributed to the degeneration of epithelia, including mucous neck cells. If Hp infection persists, the degeneration and regeneration of mucous neck cells occurs alternatively. Ultimately the generative stem cells were damaged, and as a result chronic atrophic gastritis could occur.     

Radioautographic study of binding and internalization of corticotropin-releasing factor by rat anterior pituitary corticotrophs

In order to identify the anterior pituitary cell type(s) containing corticotropin-releasing factor (CRF) receptors and to study the internalization processes of this peptide by the target cells, radioautography was performed on rat anterior pituitaries removed at specific intervals (2-60 min) after intracarotid injection of [125I]iodo-CRF into intact and adrenalectomized female rats. In intact animals, all corticotrophs were labeled, whereas in the adrenalectomized animals about 80% of the hypertrophied corticotrophs (adrenalectomy cells) were. In control animals injected with both iodinated CRF and an excess of unlabeled peptide, no specific reaction could be detected. The time-course study in intact animals showed that 2 min after injection most silver grains were found over or within 160 nm of the plasma membrane. At the 5-min time intervals, grains were observed both over the plasma membrane and within the cytoplasm, associated with lysosomes, and the Golgi apparatus. Fifteen minutes after injection, grains were mostly found over lysosomes and the Golgi apparatus, whereas at the longest time intervals (30 and 60 min) almost no labeling could be detected. The results obtained in this study indicate that in the anterior pituitary CRF receptors are restricted to corticotrophs (as identified by electron microscopy) and that, after binding to the plasma membrane, CRF is rapidly internalized to Golgi elements and lysosomes.     

Naturally occurring chronic gastritis and C pylori infection in the rhesus monkey: a potential model for gastritis in man.

Histological examination of the stomachs of Rhesus monkeys at autopsy showed chronic gastritis in a high proportion of all ages. Lesions consisted of mild to heavy infiltration of the lamina propria by lymphocytes, plasma cells, and histiocytes. The antrum was most consistently affected, but lesions were also present in the fundus and pylorus. Gastric Campylobacter-like organisms (GCLO) apparently identical to human C pylori were cultured and/or detected immunohistologically in several animals. Electron microscopy showed the spiral bacteria on the epithelial surface and in gastric pits. They did not penetrate the cells but were intimately attached to the apical plasma membrane and caused loss of microvilli. Antibodies to C pylori were detected in serum of the monkeys by ELISA. The immunospecificity of this antibody response was confirmed by Western blotting techniques. A small number of cynomolgus monkeys examined had gastritis, which may also be associated with the presence of C pylori. Baboons did not have gastritis, nor was C pylori cultured from their stomachs. The study indicates that the Rhesus monkey has a naturally occurring gastritis associated with C pylori infection and may therefore be a suitable experimental animal for the human disease.     

白水河并殖吸虫囊蚴透射电镜观察

目的观察白水河并殖吸虫囊蚴的超微形态学特征。方法应用透射电镜(TEM)观察白水河并殖吸虫囊蚴壁和囊内蚴虫。结果囊蚴壁内壁是由电子密度较低而均匀的物质构成,未见细胞和微管道结构,厚度为13.1μm,囊壁和囊内蚴虫之间有一定空隙;囊内蚴虫的体被由外皮层、肌层、皮层细胞组成,外皮层为一合体层,包括外质膜、基质、基质膜,外皮层表面呈指状突起,皮层基质内有皮棘,其根部位于基质膜,顶端被外质膜覆盖。结论通过透射电镜观察,白水河并殖吸虫囊蚴壁和囊内蚴虫的体被结构与其它几种并殖吸虫有相同点也有差异性。     

小睾并殖吸虫囊蚴和后尾蚴透射电镜观察

应用透射电镜观察小小睾并殖吸虫囊蚴和后尾蚴的超微结构,囊蚴壁是由电子密度较低而均匀的物质构成,未见细胞和微管道结构。囊内蚴虫和后尾蚴的体被由外皮层、肌层皮层细胞组成,外皮层为一合体层,包括外质膜、基质、基质膜,外皮层表面呈指状突起,后尾蚴的皮层细胞可见多条胞质小管,提示皮层是后尾蚴吸收营养物质的主要部位。