幽门螺杆菌感染对胃粘膜超微结构的影响

目的研究幽门螺杆菌（Ｈｐ）感染及其根除前后胃粘膜超微结构的变化．方法Ｈｐ感染患者１０例经三联疗法２８ｄ．Ｈｐ阴转７例于停药１月后及治疗前内镜下取胃窦粘膜，经切片染色后分别行透射电镜及扫描电镜观察．结果透射电镜显示，Ｈｐ聚集处上皮细胞微绒毛变短、减少或消失，细胞呈毛刺状或外突形成分枝状，细胞膜内侧粘液颗粒聚集，细胞破裂，释放粘液颗粒．扫描电镜下Ｈｐ横卧于微绒毛表面或垂直镶嵌在微绒毛里．应用三联疗法（德诺１２０ｍｇ＋四环素０２５ｇ＋呋喃唑酮１０ｍｇ，４次／ｄ）治疗２８ｄ，停药１月后７例Ｈｐ根除．电镜显示Ｈｐ消失，粘膜细胞变性逆转，上皮细胞及微绒毛结构恢复正常．结论Ｈｐ引起的胃粘膜超微结构损害在根除Ｈｐ后有改善及恢复     

幽门螺杆菌感染与原发性胃淋巴瘤

目的探讨幽门螺杆菌（Ｈｐ）感染与原发性胃淋巴瘤、胃ＭＡＬＴ淋巴瘤的关系．方法经内镜、手术诊断的胃淋巴瘤患者２９例，其中男１９例，女１０例；年龄３０岁～７５岁，中位数年龄５２５岁．胃窦部１３例，胃体部９例，贲门部２例．凹陷溃疡型９例，蕈样隆起型８例，结节型１２例．胃粘膜标本切片，采用ＨＥ及ＷａｒｔｈｉｎＳｔａｒｙ染色后观察组织中有无Ｈｐ感染．结果原发性胃淋巴瘤患者２９例，胃粘膜组织中发现Ｈｐ感染２４例，阳性率８２８％．其中胃ＭＡＬＴ淋巴瘤８例，Ｈｐ感染７例，阳性率８７５％．明显高于同期胃良性病变Ｈｐ阳性率４６％（Ｐ＜００１）．２９例中１１例进行了术后随访，３例仍存在Ｈｐ感染，阳性率２７３％．结论Ｈｐ感染在原发性胃淋巴瘤的发病机制中起一定作用．     

胃腺癌者Hp感染胃粘膜的电镜观察

目的：观察Ｈｐ感染胃腺癌者胃粘膜的超微结构形态特点。方法：应用透射电镜技术，对１１例经病理证实为胃腺癌者Ｈｐ感染的粘膜进行观察。结果：可见到Ｈｐ集聚于粘膜上皮绒毛陷窝，位于或接近上皮细胞连接处；主要位于粘液层下，以纤维丝样结构与上皮相连；胃粘膜表面的微绒毛减少或消失，细胞膜肿胀，不规则地形成凹陷或突起；粘液细胞发生粘液排泌、死亡，并观察细胞增殖分裂象。结论：说明Ｈｐ感染的长期存在对细胞的恶性增生有一定影响     

幽门螺杆菌感染对胃上皮细胞增殖和凋亡的影响

目的为了探讨幽门螺杆菌感染对胃粘膜上皮细胞动力学的影响。方法应用免疫组织化学和切口末端标记法（ＴＵＮＥＬ），检测了１６例正常胃粘膜者和３１例幽门螺杆菌（Ｈｐ）相关慢性胃炎患者治疗前后胃粘膜上皮细胞增殖细胞核抗原（ＰＣＮＡ）标记指数（ＬＩ％）、细胞凋亡指数（ＡＩ）和表皮生长因子受体（ＥＧＦ－Ｒ）的表达。结果Ｈｐ阳性患者的ＰＣＮＡＬＩ％为１３．９４±１．６４，正常对照组为６．７１±０．９２，差异有非常显著性（Ｐ＜０．０１）；ＥＧＦ－Ｒ表达与ＰＣＮＡＬＩ％呈正相关（ｒ＝０．４４８７，Ｐ＜０．０１）：Ｈｐ阳性患者组的ＡＩ为７．１±１．６，正常对照组为１．３±０．６，差异有非常显著性（Ｐ＜０．０１）；抗Ｈｐ治疗后，２１例Ｈｐ根除者的ＰＣＮＡＬＩ％和细胞ＡＩ分别降至８．２１±１．３２和１．２±０．６，与治疗前相比差异有非常显著性（Ｐ＜０．０１），而１０例Ｈｐ持续阳性者则无明显降低（Ｐ＞０．０５）：ＰＣＮＡＬＩ％、ＥＧＦ－Ｒ表达及细胞ＡＩ与胃粘膜炎症程度无显著相关（Ｐ＞０．０５）。结论上述结果提示，Ｈｐ感Ｈ能引起胃粘膜上皮细胞过度增殖和凋亡。这为Ｈｐ感染胃癌发病中的作用机制提供了一些线索。     

慢性胃炎患者治疗前后胃内幽门螺杆菌的分布特点

目的研究根除治疗前后幽门螺杆菌在胃内的分布特点．方法有消化道症状，Ｈｐ＋慢性胃炎患者９０例，其中男５２例，女３８例，年龄１９岁～７４岁，平均４４岁．采用ＣＢＳ，ＦｕＺ，Ｍｅｔ三联治疗２ｗｋ，在根除治疗前及治疗结束４ｗｋ后从胃底、胃体、角切迹、幽门前大弯侧多部位取活检，以ＷａｒｔｈｉｎＳｔａｒｙ染色检测Ｈｐ，ＨＥ染色观察局部胃炎程度．结果三联治疗前Ｈｐ呈全胃分布，各部位Ｈｐ阳性率无明显差异（Ｐ＞００５），但Ｈｐ数量和胃炎程度各部位有差异，且有明显相关性（ｒ＝０９４８），以角切迹Ｈｐ数量最多，胃炎程度最重，局灶性萎缩、活动性胃炎的发生率亦最高．２２例根除治疗后Ｈｐ仍阳性患者，以胃底检出率最高（９０９％），角切迹最低（３１８％），两部位相比有显著差异（χ２＝１４４，Ｐ＜００１）．结论角切迹是治疗前Ｈｐ的最佳检测部位，胃底是治疗后Ｈｐ的最佳检测部位．     

幽门螺杆菌感染患者胃氨的变化

目的探讨由Ｈｐ产生的氨在Ｈｐ致病机理中的作用．方法对１６例伴有Ｈｐ感染的消化性溃疡及慢性活动性胃炎患者，应用抗菌素对症治疗４周．分别在治疗前后用尿素酶试剂药盒检查Ｈｐ感染情况，并用Ｎｅｓｌｅｒ′ｓ显色法测定治疗前后的胃液氨浓度．结果治疗后有８例患者Ｈｐ阴转，胃氨浓度由治疗前的３０５７±１９６５μｍｏｌ／Ｌ降至１７６９±９７８μｍｏｌ／Ｌ（Ｐ＜００１），同时胃粘膜病变减轻或治愈．结论氨是Ｈｐ致病的重要因素之一．动态测定胃氨浓度可以间接反映Ｈｐ的感染情况，并可以作为判定治疗效果的一项指标．     

胃粘液层的幽门螺杆菌定植与胃炎的相关性初探

为观察胃粘液层中幽门螺杆菌（Ｈｐ）感染的情况，对２００例患者胃镜检查后检测胃粘膜及胃粘液内的Ｈｐ。结果显示，Ｈｐ不仅寄居于胃小凹及胃粘膜上皮层，还定植于胃粘液层中，且粘液中的Ｈｐ较粘膜上皮层的数量更多。局部胃炎程度不仅与粘膜上皮表面Ｈｐ密度密切相关（ｒ＝０．９９６），还与粘液层的Ｈｐ密度有较好的相关性（ｒ＝０．８２２）。提示胃粘液层Ｈｐ与胃炎相关，是否根除Ｈｐ的病理诊断应将粘液中是否存在Ｈｐ作为一项指标。     

胃癌及胃癌前病变患者中医证型与病理及Hp感染的关系

目的研究胃癌及胃癌前病变患者中医分型与病理组织学及Ｈｐ感染的关系．方法选取资料完整的慢性胃病患者之病理及临床资料２０３例进行中医分型、Ｈｐ检测和ＰＣＮＡ染色，对胃癌前病变（ｎ＝１３４）胃癌（ｎ＝３３）及对照组（浅表性胃炎，ｎ＝３６）等各组进行比较分析．结果中医分型与病理改变有一定的相关性，在浅表性胃炎中以寒热夹杂型为主，胃癌及胃癌前病变中以脾胃虚寒为主，而寒热夹杂型较少，在胃癌及胃癌前病变患者中未发现Ｈｐ感染与中医分型有明显关系，胃癌及癌前病变组Ｈｐ感染率（６２４％，６６７％～７９８％）明显高于对照组（３４１％），且发病年龄Ｈｐ阳性者明显低于Ｈｐ阴性者（４９９％ｖｓ５６７，４６１～５３８ｖｓ５３８～６２０，Ｐ＜００５），ＰＣＮＡＬＩ随病变程度的加重而升高（γ＝０９５１，Ｐ＜００２５）．结论Ｈｐ感染与胃癌及胃癌前病变的发生有密切关系．ＰＣＮＡＬＩ是反应胃粘膜病变严重程度的较客观的定量指标     

FD患者Hp感染对胃排空的影响

目的探讨幽门螺杆菌（Ｈｐ）感染在功能性消化不良（ＦＤ）患者胃排空功能障碍中的作用．方法采用放射性同位素γ－照相法观测了５６例ＦＤ患者胃固－液体排空情况，并用胃窦粘膜印片Ｇｉｅｍｓａ染色及石蜡切片ＨＥ，Ｗ＿Ｓ银染色镜检Ｈｐ．结果ＦＤ患者餐后３０，６０及９０ｍｉｎ时的胃排空率均显著低于正常对照组（Ｐ＜００５－００１）；ＦＤ患者Ｈｐ感染率无明显增高（Ｐ＞００５），Ｈｐ阳性组与阴性组在３个时相的胃排空率差异均无显著性（Ｐ＞００５）结论ＦＤ患者胃排空功能与Ｈｐ感染无关     

消化性溃疡患者Hp感染与粘膜血流量

目的研究幽门螺杆菌（Ｈｐ）感染与消化性溃疡（ＰＵ）胃肠粘膜血流量（ＧＤＭＢＦ）的关系．福建省立医院消化内科福建省福州市３５０００１方法采用多普勒激光血流仪测定内镜检查患者的非病灶区贲门、胃体、胃窦部及溃疡或胃炎病灶区的ＧＤＭＢＦ．结果非病灶区ＧＤＭＢＦ以贲门部最高，胃体部次之，胃窦部较低，分别为１．６８±０５５Ｖ，１７０±０４２Ｖ和１３５±０３７Ｖ，均明显高于ＰＵ病灶区０８９±０３３Ｖ（Ｐ＜００１）；伴有Ｈｐ感染的ＰＵ患者病灶区及其它非病灶区ＧＤＭＢＦ均显著低于不伴Ｈｐ感染者．结论Ｈｐ感染可造成胃肠粘膜血流量降低，可能是溃疡病灶难愈或复发的原因之一．     

Appraisal of polyethyleneimine used as a tracer for anionic sites in human cardiac tissues with a brief reference to anionic molecular organization in altered basement membranes

Electron microscopic cytochemical studies on human ventricular muscles obtained during open heart surgery were carried out in ten patients with tetralogy of Fallot. Polyethyleneimine was used as a tracer for demonstration of anionic sites in the membrane surfaces of myocardial cells and capillaries. In the myocardial cells PEI particles were consistently observed to be orderly arranged in the external lamina of normal basement membranes at regular spacings of 40 to 80 nm. Few PEI particles were irregularly dispersed in the surface coat of basement membranes and the interspaces of intercalated discs. PEI particles were also seen to be regularly distributed in normal capillary basement membranes, mainly restricted in outer lamina densa at intervals of 40 to 80 nm. It was of particular interest that PEI particles were often seen to be irregularly and loosely arranged in abnormally thickened basement membranes of myocardial cells and capillaries. In addition, focal loss of PEI deposition in altered basement membranes was a frequent finding. Based on electron microscopic cytochemical findings the following conclusions are made: The anionic sites characterized by PEI which is considered to be superior to most polycationic colloids show a regular lattice-like arrangement in the basement membranes of human myocardial cells and capillaries; Anionic groups which are not distributed uniformly on human myocardial cell surfaces show different distribution patterns in the external lamina and surface coat of basement membranes; and Perturbations of regular anionic arrays are demonstrable in altered basement membranes of diseased human myocardium.     

肥大心肌细胞膜阴离子位点的变化

目的 :旨在研究肥大心肌细胞膜损伤的分子病理学基础。方法 :本实验采用大鼠腹主动脉缩窄模型 ,以聚乙烯亚胺 (PEI)为阳离子探针 ,观察肥大心肌细胞膜基板阴离子位点的变化 ,并测定心肌细胞内的Ca2 + 含量和细胞膜的Ca2 + ATP酶活性。结果 :对照组大鼠心肌细胞表面PEI颗粒沿基膜以 4 0～ 80nm间距呈较规则的点阵式、线性排列。手术组心肌细胞表面PEI颗粒明显减少 ,并可见心肌细胞基膜与质膜分离 ,基膜断裂 ;心肌细胞内的Ca2 + 含量增加 ,细胞膜的Ca2 + ATP酶活性降低。结论 :表明肥大心肌细胞膜基板阴离子位点减少 ,膜保护屏障功能下降     

Distribution and movement of anionic cell surface sites in cultured human vascular endothelial cells.

Cell surface anionic sites on primary and transformed cultures of human vascular endothelium were studied using cationized ferritin (CF) as an ultrastructural marker. The native distribution of anionic sites on the upper (free) surfaces of cells, fixed in situ with glutaraldehyde, was uniform. Binding of the polycationic ligand, CF, in unfixed cells induced redistribution of anionic sites. Rapid formation of discrete patches of CF particles was followed by reappearance of binding between patches, movement of surface-bound CF into intercellular clefts, and endocytosis, over the next 30 min. Aldehyde-fixed cells, detached from the culture surface, bound CF on both upper and lower surfaces. The distribution and mobility of negatively charged membrane components in vascular endothelium may have relevance for thrombosis, atherogenesis, and vascular permeability.     

Distribution of cell surface charges on mesothelium and lymphatic endothelium

The distribution of anionic sites on the luminal surfaces of the peritoneal mesothelium and lymphatic endothelium was investigated by injecting cationized ferritin (CF) intraperitoneally. After washing with phosphate-buffered saline, the diaphragm was fixed and processed for electron microscopy. CF label occurred in discontinuous patches along the mesothelial surface. Microvilli were heavily marked and often closely applied to the mesothelial surface. The intercellular cleft was also heavily labeled. The luminal aspect of the lymphatic endothelium was more extensively labeled, with the marker occurring in long discontinuous dense bands. The clefts of lymphatic endothelial intercellular junctions were extensively labeled especially along regions where cells were loosely apposed. The existence of a high density of anionic sites on membranes at the intercellular junctions of both mesothelial and lymphatic endothelial cells represent a salient feature which is very different from binding in blood capillary endothelium. The presence of a high density of anionic sites along the intercellular clefts of adjacent cells may play a role in the rapid movement of small solutes and molecules from interstitial spaces into the lymphatic lumen.     

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.     

Alterations of glomerular basement membrane charge and structure in diabetic nephropathy

Summary We examined glomerular basement membrane anionic site distribution identified by cationic gold in seven patients with insulin-dependent and four patients with non-insulin-dependent diabetes mellitus, presenting a spectrum of clinical and glomerular changes. Anionic sites were investigated by pretreatment of tissue with glycosaminoglycan-degrading enzymes prior to cationic gold staining. The distribution of chondroitin sulphate proteoglycans — a previously unrecognized glomerular basement membrane component — and type IV collagen was examined by immunoelectron microscopy to identify structural changes in the basement membrane. Findings were compared with those of non-diabetic patients showing minor proteinuria and morphologically normal glomerular basement membranes. Two patients, originally diagnosed as having diabetic nephropathy were also examined at 19 weeks and 5 years after renal transplantation. Characteristic redistribution of type IV collagen and chondroitin sulphate proteoglycans was noted in thickened glomerular basement membrane segments (>400 nm) of diabetic patients and those with renal transplants. Extension of anionic sites deep into the glomerular basement membrane at pH 2.5, together with loss of interna sites at pH 5.8 is unique to diabetic nephropathy. Reduced charge density was apparent in some patients due to thickening of the glomerular basement membrane, although the number of anionic sites per unit length of membrane was actually increased. Thus, charge aberration in diabetic nephropathy is due to displacement rather than loss of anionic sites. Removal of more than 90% of these sites by heparitinase, confirms their association with heparan sulphate proteoglycans. Similar derangement of anionic sites in all patients with diabetic nephropathy irrespective of the degree of proteinuria, suggests that a heparan sulphate proteoglycan-related charge barrier plays a minor role in controlling permeability of the diabetic glomerular basement membrane.Abbreviations BSA Bovine serum albumin - CG cationic gold - CSPG chondroitin sulphate proteoglycans - GAG glycosaminoglycan - GBM glomerular basement membrane - HSPG heparan sulphate proteoglycans - LRE lamina rara externa - LRI lamina rara interna - PCI protein:creatinine index     

Reduction of heparan sulphate-associated anionic sites in the glomerular basement membrane of rats with streptozotocin-induced diabetic nephropathy

Summary Heparan sulphate-associated anionic sites in the glomerular basement membrane were studied in rats 8 months after induction of diabetes by streptozotocin and in age- and sex-matched control rats, employing the cationic dye cuprolinic blue. Morphometric analysis at the ultrastructural level was performed using a computerized image processor. The heparan sulphate specificity of the cuprolinic blue staining was demonstrated by glycosaminoglycan-degrading enzymes, showing that pretreatment of the sections with heparitinase abolished all staining, whereas chondroitinase ABC had no effect. The majority of anionic sites (74% in diabetic and 81% in control rats) were found within the lamina rara externa of the glomerular basement membrane. A minority of anionic sites were scattered throughout the lamina densa and lamina rara interna, and were significantly smaller than those in the lamina rara externa of the glomerular basement membrane (p<0.001 and p<0.01 for diabetic and control rats, respectively). Diabetic rats progressively developed albuminuria reaching 40.3 (32.2–62.0) mg/24 h after 8 months in contrast to the control animals (0.8 (0.2–0.9) mg/24 h, p<0.002). At the same time, the number of heparan sulphate anionic sites and the total anionic site surface (number of anionic sites × mean anionic site surface) in the lamina rara externa of the glomerular basement membrane was reduced by 19% (p<0.021) and by 26% (p<0.02), respectively. Number and total anionic site surface in the remaining part of the glomerular basement membrane (lamina densa and lamina rara interna) were not significantly changed. We conclude that in streptozotocin-diabetic rats with an increased urinary albumin excretion, a reduced heparan sulphate charge barrier/density is found at the lamina rara externa of the glomerular basement membrane.Abbreviations HS Heparan sulphate - GBM glomerular basement membrane - HSPG heparan sulphate proteoglycan - STZ streptozotocin - LRE lamina rara externa - LD lamina densa - LD + LRI lamina densa + lamina rara interna - ANOVA analysis of variance     

Anionic sites in basement membranes. Differences in their electrostatic properties in continuous and fenestrated capillaries

We have used ruthenium red, a cationic dye, to detect at the electron microscopic level the presence of anionic sites in various murine basement membranes, with particular emphasis on those of the microvasculature. We have observed anionic sites in all continuous and fenestrated capillaries examined. Terminal lymphatics, which have a discontinuous basement membrane, have sites only where the basement membrane is present. Anionic sites are not present beneath sinusoidal lining cells of the liver which lack a basement membrane. Basement membranes of epithelial cells and those surrounding striated and smooth muscle cells, pericytes, fat cells, and Schwann cells also exhibit anionic sites. We compared the electrostatic properties of anionic sites in basement membranes of continuous and fenestrated capillaries by determining the salt concentration (critical electrolyte concentration, Scott and Dorling, 1965) required to displace ruthenium red from the sites. A concentration of 0.5 M Na⁺ was required to displace ruthenium red from the basement membrane of continuous capillaries of muscle whereas 1.3 M Na⁺ was required to displace ruthenium red from the basement membrane of fenestrated peritubular capillaries of the renal cortex. Our results suggest that anionic sites in the basement membrane of fenestrated peritubular capillaries are more strongly negatively charged than those in the basement membrane of continuous capillaries of muscle. We conclude from this study, first, that anionic sites are a general property of vascular, epithelial, and pericellular basement membranes and, second, that the electrostatic properties of the sites differ in different vascular basement membranes. We speculate that the anionic sites in vascular basement membranes and the variation in their electrostatic properties in different types of capillaries may have important implications for exchange of substances across the capillary wall.     

Hp与慢性胃炎和消化性溃疡粘膜上皮关系的超微结构研究

目的应用透射电子显微镜(TEM)研究幽门螺杆菌(Hp)与慢性胃炎和消化性溃疡粘膜上皮的关系.方法 75例患者胃窦部活检标本用戊二醛固定,在锇酸染色之前先用单宁酸处理,常规包埋,TEM 观察(每份样品至少做4张半薄切片定位,再行超薄切片).受检患者接受6种其它检测 Hp 方法试验.结果 55例感染上 Hp 的患者中有53例胃粘膜内有 TEM 检查到细菌.在单宁酸特别染色的超薄切片中清楚地显示 Hp 通过糖萼与胃粘膜上皮相连.当 Hp 呈克隆状生长繁殖时,邻近的粘液细胞以及颈粘液细胞变性,其特征是细胞朝腺腔的胞质部分溃烂,细胞空泡样变或大泡形成,甚至整个上皮脱落.综合许多的证据表明:位于腺腔Hp吸引中性粒细胞迁移到上皮细胞的间隙中,或者进入腺腔内进行吞噬 Hp 活动.结论用 TEM 诊断 Hp 感染的敏感和特异性分别是96%和95%.单宁酸能较好地保存细胞的糖萼.呈克隆状生长、多带有质周池的 Hp 是导致粘膜上皮,包括颈粘液细胞,发生一系列变性的原因.若 Hp 感染持续存在,颈粘液细胞变性和再生交替进行.最后再生干细胞受损伤,结果导致慢性萎缩性胃炎发生.