我国山丘疫区和平原疫区利什曼原虫分离株基因组DNA基因型分析

目的：分析我国山丘和平原疫区利什曼原虫分离株基因组DNA（nDNA）的多态性。方法：对基因组DNA进行内切酶酶切、Southern杂交、染色体定位。探针采用地高辛标记。结果：采用gp63探针，显示杜氏利什曼原虫（Leishmaniadonovani，L．d．）江苏人株和L．d．Jeddah nDNA之间有相似的染色体杂交带，L．d．四川人株和L．infantum nDNA之间有相似的染色体杂交带；采用β－tubulin探针，显示L．d．江苏人株与L．d．Jeddah nDNA之间有两条相似的染色体杂交带，L．d．四川犬株与L．d．甘肃犬株nDNA之间有三条相似的染色体杂交带、与L．d．汶川人株nDNA之间有两条相似的染色体杂交带。结论：提示平原疫区的江苏人株与L. d. J eddah 之间存在同源性, 山丘疫区的L. d. 四川人株与L. infantum 之间存在同源性, L. d. 四川犬株与L. d. 甘肃犬株之间存在同源性、与L. d. 汶川人株之间既存在同源性又存在差异, 山丘疫区与平原疫区分离株之间存在异源性。     

细胞因子体外扩增脐血CD^＋34细胞移植SCID鼠人类免疫功能的 …

ObjectiveTo explore the hematopoietic function of cytokine-expanding CD⁺₃₄ cells from umbilical cord blood in vitro.MethodCord blood cells expanded with FLT3(FMS-like tyrosine kinase 3)-ligand and thrombopoietin were transplanted in SCID(severe combined immune-deficient)mice. Human mature cell surface markers were detected with flow cytometry and used as indicator of succeeding transplantation.ResultAfter 2 and 3 weeks culture,the number of CD⁺₃₄ cells increased approximately 10 and 37 folds. After transplantation with 6.6×10⁴ to 6.9×10⁵ CD⁺₃₄ cells,respectively the percentage of human CD⁺₄₅ cells in the bone marrow of all 10 SCID mice is 0.2%～4.0%. The percentage of human CD⁺₃ cells in the peripheral blood and spleen of 6 SCID mice is 0.1%～2.3%. The percentage of human CD⁺₁₉ cells in the peripheral blood and spleen of 3 SCID mice are 0.3%～1.3%.ConclusionCD⁺₃₄ cells expanded with FLT3-ligand and thrombopoietin are potential engraft used for reconstitution of both hematopoietic and immunologic function of SCID mice.     

Catheter ablation of atrial fibrillation in elderly population

Background Although elderly patients have been included in published series of catheter ablation for atrial fibrillation (AF), clinical benefit and safety remain still less defined in this population. A retrospective analysis of the results of catheter ablation for AF in a large volume center focused on comparison of elderly patients with the rest of the patient cohort was conducted in this study. Methods Consecutive patients who underwent catheter ablation for AF between January 2001 and December 2016 were analysed. A total population of 3197 patients was dichotomized by the age of 70 years (394 elderly vs. 2803 younger subjects). Patients were followed in terms of arrhythmia status and sur?vival for a median period of 18 vs. 21 and 35 vs. 57 months, respectively. Results Elderly patients were more frequently females (49% vs. 29%, P < 0.0001), had a history of hypertension (79% vs. 57%, P < 0.0001), diabetes (16% vs. 11%, P < 0.01), stroke (9% vs. 6%, P < 0.01), coronary/peripheral artery disease (14% vs. 8%, P < 0.0001), and CHA2DS2-VASc score (3.1 ± 1.3 vs. 1.5 ± 1.2 s, P < 0.0001). Major complications were more frequent in elderly (5.3% vs. 3.2%, P = 0.03); however, this difference was driven by vascular complications (3.6% vs. 1.9%, P = 0.04). There were comparable rates of cerebrovascular (0.3 vs. 0.3%) or nonvascular complications (1.8 vs. 1.2%). Good arrhythmia control was inferior in elderly patients as compared with the rest of the cohort, both without and with antiarrhythmic drugs: 44.2% vs. 58.2% (P < 0.0001) and 78.2 vs. 83.2% (P < 0.01), respectively. Poor arrhythmia control was associated with relative risk of all-cause mortality of 2.7 (95% CI: 1.1–6.4) in elderly patients and 1.4 (95% CI: 0.9–2.0) in younger subjects. Conclusions Catheter abla?tion for AF in elderly patients is safe although somewhat less effective. Good arrhythmia control is associated with better survival, especially in elderly patients.     

Comparison of in-hospital outcomes between octogenarians and nonagenarians undergoing transcatheter aortic valve replacement: a propensity matched analysis

Background Aortic valve stenosis (AS) is very common in the elderly patients above 80 years. Transcatheter aortic valve replacement (TAVR) in such patients is being increasingly performed. This study sought to assess in-hospital outcome differences between octogenarians and nonagenarians and predictors of mortality in nonagenarians undergoing TAVR with severe AS. Method The study population was derived from the National Inpatient Sample (NIS) for the years 2012–2014 using ICD-9 CM procedure codes 35.05 and 35.06 for TAVR. Hospitalizations below 80 years of age were excluded. After performing propensity score matching (1: 2), in-hospital outcomes were compared in matched cohorts. Then, multivariate model was developed to analyze predictors of in-hospital mortality in nonagenarians. Results There were 11,630 hospitalizations in the octogenarian and 5815 hospitalizations in the nonagenarian group. Primary outcome of in-hospital mortality (6% vs. 4.1%, P ≤ 0.001) was higher in nonagenarians compared to octogenarians. Secondary outcomes including stroke (3.4% vs. 2.8%, P ≤ 0.001), renal failure (18.9% vs. 17.3%, P ≤ 0.001), blood transfusion (35% vs. 32.6%, P ≤ 0.001), vascular complications (4.5% vs. 3.5%, P ≤ 0.001), and pacemaker implantation (27.8% vs. 24.8%, P ≤ 0.001) were higher in nonagenarians. There was no difference in their length of stay. Median cost (70,374$ vs. 65,381$, P ≤ 0.001) was slightly higher with nonagenarian. Conclusions Although in-hospital mortality is slightly higher in nonagenarians, it is acceptable. This difference in mortality is at least partly explained by higher complications in nonagenarians. Efforts should be made to decrease the complications which can further narrow the difference in in-hospital mortality between the groups.     

The comparative efficacy of ezetimibe added to atorvastatin 10 mg versus uptitration to atorvastatin 40 mg in subgroups of patients aged 65 to 74 years or greater than or equal to 75 years

Background Coronary heart disease (CHD) risk increases with age; yet lipid-lowering therapies are significantly under-utilized in patients > 65 years. The objective was to evaluate the safety and efficacy of lipid-lowering therapies in older patients treated with atorvastatin 10 mg + ezetimibe 10 mg (EZ/Atorva) vs. increasing the atorvastatin dose to 40 mg. Methods Patients ≥ 65 years with atherosclerotic vascular disease (LDL-C ≥ 1.81 mmol/L) or at high risk for coronary heart disease (LDL-C ≥ 2.59 mmol/L) were randomized to EZ/Atorva for 12 wk vs. uptitration to atorvastatin 20 mg for 6 wk followed by atorvastatin 40 mg for 6 wk. The percent change in LDL-C and other lipid parameters and percent patients achieving prespecified LDL-C levels were assessed after 12 wk. Results EZ/Atorva produced greater reductions in most lipid parameters vs. uptitration of atorvastatin in patients ≥ 75 years (n = 228), generally consistent with patients 65–74 years (n = 812). More patients achieved LDL-C targets with combination therapy vs. monotherapy in both age groups at 6 wk and in patients ≥ 75 years at 12 wk. At 12 wk, more patients ≥ 75 years achieved LDL-C targets with monotherapy vs. combination therapy. EZ/Atorva produced more favorable improvements in most lipids vs. doubling or quadrupling the atorvastatin dose in patients ≥ 75 years, generally consistent with the findings in patients 65–74 years. Conclusions Our results extended previous findings demonstrating that ezetimibe added to a statin provided a generally well-tolerated therapeutic option for improving the lipid profile in patients 65 to 74 years and ≥ 75 years of age.     

Growth hormone-releasing hormone and gonadotropin-releasing hormone stimulate nitric oxide production in 17beta-estradiol-primed rat anterior pituitary cells

It was reported that neuronal nitric oxide synthase (nNOS) was expressed only in gonadotrophs and folliculo-stellate cells in the anterior lobe of the pituitary gland. However, recent studies have demonstrated the occurrence of nNOS in the somatotrophs and lactotrophs. In the present study, we investigated effects of growth hormone-releasing hormone (GHRH), gonadotropin-releasing hormone (GnRH), and 17β-estradiol on nitric oxide (NO) release in cultured rat anterior pituitary cells in vitro. The NO ₂ ⁻ level in the incubation medium of the rat anterior pituitary cells was dependent on the cell density. Pretreatment with 10 μM 17β-estradiol resulted in an increase in medium NO ₂ ⁻ level. GHRH and GnRH failed to change medium NO ₂ ⁻ levels, but they elicited increases in medium NO ₂ ⁻ levels in estrogen-treated cells. The GHRH-induced increase in NO ₂ ⁻ level was inhibited by Nχ-nitro-l-arginine methyl ester, a NOS inhibitor. These findings suggest that GnRH and GHRH could activate nNOS in the gonadotrophs and the somatotrophs, respectively.     

我国伊蚊族蚊类记录的校订及新分类系统的建议（双翅目∶蚊科）

本文根据国外蚊虫分类经验,对我国伊蚊族蚊类以往记录171种的分类地位作了校订,其中160种可归入我国伊蚊族新分类系统的29个属中,另外11种地位未定,暂按旧分类地位列名表供查考。我国伊蚊族新分类系统包括伊蚊属Aedes、阿蚊属Armigeres、艾蚊属Ayurakitia、博蚊属Bothaella*、布蚊属Bruceharrisonius*、环喙蚊属Christophersiomyia*、科蚊属Collessius*、丹蚊属Danielsia*、唐蚊属Downsiomyia*、箭阳蚊属Edwardsaedes*、纷蚊属Finlaya*、弗蚊属Fredwardsius*、贾蚊属Gilesius*、领蚊属Heizmannia、喜蚊属Himalaius*、霍金蚊属Hopkinsius*、呼蚊属Hulecoeteomyia*、连蚊属Jihlienius*、奈蚊属Kenknightia*、陆蚊属Luius*、霉蚊属Mucidus*、新黑蚊属Neomelaniconion*、骚扰蚊属Ochlerotatus、花蚊属Phagomyia*、盾蚊属Scutomyia*、覆蚊属Stegomyia*、田中蚊属Tanakaius*、尤蚊属Udaya和奇阳蚊属Verrallina等29属,其中22属（*）为我国新记录。此外,还对《中国动物志,昆虫纲第8卷,双翅目：蚊科上卷》记载的4种伊蚊作了重要订正。安图伊蚊Ae. （Edw.） antuensis应是平坝箭阳蚊Ed. pingpaensis的同物异名。滇西伊蚊Ae. （Sin.） occidentayunnanus、黄背伊蚊Ae. （Och.） flavidorsalis和亚同伊蚊Ae. （Fin.） subsimilis分别订正为滇西领蚊Hz. （Mat.） occidentayunnana、白色骚扰蚊Oc. albineus和亚同尤蚊Ud. subsimilis。     

Formoterol,a new long-acting selectiveβ 2-agonist,decreases airway responsiveness in children with asthma

We compared the protective effect and duration of action of inhaled formoterol with salbutamol and placebo in 16 asthmatic children in a double-blind, cross-over study. All had an FEV₁ ≥ 70% predicted normal and a provocative concentration of methacholine (MCh) required to decrease their FEV₁ by 20% (PC₂₀) ≤ 4 mg/ml. On each study day, FEV₁ was within 10% and PC₂₀ within one doubling-dose of the initial visit. Patients received either placebo, salbutamol 200μg, formoterol 12μg, or formoterol 24μg by metered-dose inhaler. FEV₁ and PC₂₀ were measured repeatedly over 12 h. After salbutamol, peak FEV₁ was 120% of baseline at 30 min and returned to baseline in 3 h. After formoterol (12 or 24μg) peak FEV₁ was 118% at 3 h and remained above baseline for at least 12 h. Protection from MCh by both doses of formoterol was significantly better than by salbutamol. Protection from formoterol 12 and 24μg at 12 h was equivalent to that from salbutamol at 3 h. The PC₂₀ of four children 48 h after formoterol 24μg was more than twice their baseline PC₂₀. Formoterol by inhalation is potent and long-acting and provides significantly better antiasthma protection than salbutamol.     

Real-world comparison of non-vitamin K antagonist oral anticoagulants and warfarin in Asian octogenarian patients with atrial fibrillation

Background The efficacy and safety of non-vitamin K antagonist oral anticoagulants (NOACs) and warfarin in Asian octogenarian atrial fibrillation (AF) patients have not been established in a real-world setting. We aimed to evaluate the efficacy and safety of NOACs and warfarin in Korean octogenarian patients. Methods A total of 293 consecutive patients aged ≥ 80 years with non-valvular AF who had taken either NOACs (148 cases, 50.5%) or warfarin (145 cases, 49.5%) were retrospectively reviewed. The efficacy outcome was the composite of stroke or systemic embolism. The safety outcome was major bleeding. Results The follow-up duration was 375 patient-years (172 patient-years with NOACs and 203 patient-years with warfarin). Patients on NOACs were slightly older (P = 0.006) and had slightly higher HAS-BLED scores (P = 0.034). The efficacy of both anticoagulants was high (1.16% for NOACs vs. 2.98% for warfarin per 100 patient-years, P = 0.46). The safety outcome was relatively high in both NOACs and warfarin groups (8.96% vs. 12.46%, P = 0.29). The efficacy and safety outcomes tended to decrease non-significantly in low dose NOACs than in common dose NOACs or warfarin (0.85% vs. 1.84% vs. 2.98% in efficacy outcome, P = 0.69; and 6.97% vs. 13.29% vs. 12.46% in safety outcome, P = 0.34). Conclusions NOACs were highly effective for prevention of stroke or systemic embolism in Asian octogenarian AF patients. However, major bleeding occurred excessively high in both anticoagulant groups. Further study is required on the optimal anticoagulant regimen in octogenarian population.     

Expression of nitric oxide synthases and effects of L-arginine and L-NMMA on nitric oxide production and fluid transport in collagenous colitis

BACKGROUND AND AIMS: Luminal nitric oxide (NO) is greatly increased in the colon of patients with collagenous and ulcerative colitis. To define the source and consequence of enhanced NO production we have studied expression of NO synthase (NOS) isoforms and nitrotyrosine in mucosal biopsies from these patients. In addition, effects on colonic fluid transfer caused by manipulating the substrate of NOS were studied in patients with collagenous colitis. PATIENTS: Eight patients with collagenous colitis, nine with active ulcerative colitis, and 10 with uninflamed bowel were included. METHODS: Expression of NOS isoforms was quantified by western blotting. Inducible NOS (iNOS) and nitrotyrosine were localised by immunohistochemistry. Modulation of NOS activity by topical N(G)-monomethyl-L-arginine (L-NMMA) or L-arginine was assessed during perfusion of whole colon. Plasma and perfusate nitrite/nitrate (NOx) was measured by Griess' reaction. RESULTS: Both in collagenous and ulcerative colitis, expression of iNOS was 10(2)-10(3) higher (p<0.001) than in uninflamed bowel and localised primarily to the epithelium. Endothelial NOS was evenly expressed in all groups while neuronal NOS was undetectable. Nitrotyrosine was markedly expressed in active ulcerative colitis but rarely detected in collagenous colitis and never in uninflamed bowel. In collagenous colitis, the output of NOx was markedly increased compared with uninflamed bowel (283 (58) v <37 nmol/min; p<0.01) and fluid was net secreted. L-NMMA reduced the output of NOx by 13-66% (95% confidence intervals) and secretion of fluid by 25-109% whereas L-arginine increased the output of NOx by 3-39% and secretion of fluid by 15-93%. CONCLUSIONS: In collagenous colitis, as opposed to ulcerative colitis, upregulation of iNOS occurs in the absence of nitrotyrosine formation and mucosal damage. Excess generation of NO may be the primary cause of diarrhoea in this condition.     

Intestinal epithelial cells contribute to the enhanced generation of platelet activating factor in ulcerative colitis.

Generation of platelet activating factor by intestinal mucosal epithelial cells and lamina propria mononuclear cells was evaluated to elucidate the possible role of this mediator in the pathogenesis of inflammatory bowel disease. Epithelial and lamina propria mononuclear cells were isolated from surgical specimens from control, Crohn''s disease, and ulcerative colitis patients. Platelet activating factor was extracted from highly purified cell preparations with 80% ethanol after stimulation with and without 0.2 uM calcium ionophore A23187 and was measured by platelet aggregation assay. Both cell types generated platelet activating factor activity and this was generally comparable for epithelial and lamina propria cells. Basal and stimulated platelet activating factor activity of epithelial and lamina propria cells from ulcerative colitis but not Crohn''s disease patients was appreciably higher than that of control. Stimulation with calcium ionophore increased appreciably platelet activating factor activity in lamina propria cells from all groups. In contrast, only epithelial cells from ulcerative colitis showed an appreciable increase after calcium ionophore induction. These results suggest that epithelial cells are important contributors to intestinal platelet activating factor generation under normal and inflammatory conditions and that epithelial cells actively play a part in the pathogenesis of ulcerative colitis.     

Lymphocytic infiltration and expression of inducible nitric oxide synthase in human duodenal and colonic mucosa is a characteristic feature of ankylosing spondylitis

OBJECTIVE: In patients with ankylosing spondylitis (AS), inflammatory processes have been detected in the ileal and colonic mucosa. The inducible isoform of nitric oxide synthase (iNOS) may be expressed early in the inflammatory process. We investigated iNOS activity and lymphocytic infiltration in the duodenum and colon in patients with AS and ulcerative colitis compared with controls. METHODS: Gastroscopy with duodenal biopsies and/or colonoscopy with biopsies were conducted in 42 patients with AS treated or not treated with nonsteroidal antiinflammatory drugs (NSAID), in 15 with ulcerative colitis, and in 46 controls. Lymphocytic infiltration in the lamina propria and intraepithelial infiltration were quantified by histological score. iNOS expression was assessed by immunohistochemistry with monoclonal antibodies, and iNOS activity was determined by radiochemical assay. RESULTS: Endoscopic examination of the gastroduodenal or colonic mucosa did not reveal macroscopic lesions in the AS patients. In the duodenum, mucosal lymphocytic infiltration was found in 83.3% of the AS group compared to 48.6% of controls (p = 0.02), and was independent of the NSAID intake. Intraepithelial lymphocyte infiltration was increased in both duodenum and colon in AS patients compared to controls. iNOS activity in duodenum and colon and expression of iNOS protein in lamina propria inflammatory cells was increased in AS patients compared to controls. CONCLUSION: Lymphocytic infiltration and iNOS expression and activity were detected in duodenal and colonic mucosa from patients with AS. Such findings may indicate an inflammatory process in the small intestine and colon of patients with AS.     

Suppression of pro-inflammatory cytokine release by selective inhibition of inducible nitric oxide synthase in mucosal explants from patients with ulcerative colitis

BACKGROUND: In ulcerative colitis (UC), inflammatory damage is associated with increased production of pro-inflammatory cytokines and nitric oxide through the inducible nitric oxide synthase (iNOS) pathway. In an animal model of acute experimental colitis we have previously shown amelioration of inflammation with the highly selective iNOS inhibitor 1400W. The aim of the present study was to investigate the effects of selective iNOS inhibition on the production of pro-inflammatory cytokines by the colon mucosa in UC. METHODS: Inflamed and uninflamed mucosa from patients with severe UC were incubated with a highly selective iNOS inhibitor N-[3-(aminomethyl)benzyl]acetamidine (1400W), with a relatively selective cNOS inhibitor N(G)-nitro-L-arginine-methyl-esther (L-NAME), or with an NO-donor, S-nitroso-acetylpenicillamine (SNAP). Cytokine concentrations in the incubation medium were quantitated with ELISA. RESULTS: Compared to uninflamed mucosa there was an increase in iNOS protein and nitrotyrosine levels in inflamed mucosal samples. Immunolocalization of iNOS and nitrotyrosine showed their expression in inflammatory cells in the lamina propria. Expression of iNOS was also found in the epithelial brush border. Selective inhibition of iNOS suppressed the release of tumour necrosis factor alpha (TNF-alpha, by 66%) and interleukin-6 (IL-6, by 27%). The NO-donor, SNAP, augmented the secretion of TNF-alpha, IL-6 and IL-1-beta (by 62%, 52% and 175%, respectively) and decreased the release of IL-1 receptor antagonist (IL-1Ra, by 34%) by the inflamed mucosa. Moreover, in uninflamed samples, 1400W suppressed the production of TNF-alpha (by 69%) and incubation with SNAP decreased IL-6 concentrations by 48%. The cNOS over iNOS selective inhibitor L-NAME had no significant effects on the accumulation of cytokines. CONCLUSION: Selective inhibition of iNOS suppresses mucosal TNF-alpha and IL-6 release in active UC, whereas NO seems to exacerbate the inflammatory response. These results suggest that selective iNOS inhibition may have therapeutic promise in the treatment of UC.     

Phenotypes and spontaneous immunoglobulin production in mononuclear cells suspensions isolated from colonic biopsies of patients with mild active and quiescent ulcerative colitis

Lamina propria mononuclear cells can be isolated from mucosal specimens of human colon. In the present study, we have explored whether both the phenotypes and functional properties can be studied in lamina propria mononuclear cell suspensions isolated from the same set of endoscopic biopsies in patients with ulcerative colitis. The counts of CD11b+ lamina propria mononuclear cells in mild active ulcerative colitis were significantly higher than those of both quiescent ulcerative colitis and controls. Similarly, the CD16+ and the CD19+ lamina propria mononuclear cells were significantly increased in mild ulcerative colitis patients in comparison to both quiescent ulcerative colitis and control lamina propria mononuclear cells. Lamina propria mononuclear cells from all the biopsy samples appeared to produce detectable amounts of immunoglobulins of the three classes. The production of IgG in mild ulcerative colitis cultures was significantly higher than that observed in quiescent ulcerative colitis and controls. In contrast, the production of IgA in active ulcerative colitis lamina propria mononuclear cell cultures appeared to be significantly lower than that of both quiescent ulcerative colitis and controls. This study shows that morphology, phenotypes, and functional properties can be assessed in lamina propria mononuclear cell suspensions obtained from the same set of endoscopic biopsy samples. We have also shown that changes in phenotypes and functional status of lamina propria mononuclear cells occurred in mild active ulcerative colitis while no significant abnormality of these parameters was found in quiescent ulcerative colitis. This indicates that a normalization of mucosal immune functions occurs in ulcerative colitis patients when complete clinical and histological remission is achieved.     

骨桥蛋白在炎症性肠病患者结肠组织中的表达

背景：骨桥蛋白（OPN）是一种分泌型磷酸化糖蛋白,参与细胞信号转导,促进细胞黏附和迁移。近年研究发现炎症性肠病（IBD）患者血浆OPN水平升高,患者结肠黏膜中可检出OPN表达。目的：了解OPN在IBD患者结肠黏膜组织中的表达情况,探讨其在IBD发病机制中的作用。方法：以免疫组化半定量方法检测53例溃疡性结肠炎（UC）、62例克罗恩病（CD）和15例源自结肠腺瘤患者的正常结肠组织标本中OPN的表达。结果：OPN广泛表达于结肠黏膜上皮细胞和固有层渗出细胞。UC组和CD组结肠上皮细胞的OPN平均光密度值显著低于对照组（11．84±0．98和10．04±1．37对12．64±1．45,P〈0．05）,结肠黏膜固有层渗出细胞的OPN阳性细胞百分率则显著高于对照组（20．31％±4．50％和12．69％±3．06％对3．85％±0．66％,P〈0．001）。UC和CD患者的病情严重程度与结肠上皮细胞和固有层渗出细胞中的OPN表达量均无相关性。结论：OPN在IBD患者结肠黏膜上皮和同有层中的表达不一致,结肠上皮细胞中OPN表达下调可能与肠黏膜屏障功能受损有关,而黏膜固有层渗出细胞中OPN表达上调可能与免疫反应有关。     

Location of tumour necrosis factor alpha by immunohistochemistry in chronic inflammatory bowel disease.

This study determined the location and tissue density of cells immunoreactive for tumour necrosis factor alpha (TNF alpha) in intestinal specimens from 24 patients with chronic inflammatory bowel disease (15 with Crohn's disease, nine with ulcerative colitis) and 11 controls. There was significantly increased density of TNF alpha immunoreactive cells in the lamina propria of both ulcerative colitis and Crohn's disease specimens, although the distribution of these cells differed in the two conditions. In ulcerative colitis most of the TNF alpha immunoreactivity was seen in the subepithelial macrophages, with comparatively less in the deep lamina propria, while in Crohn's disease immunoreactive cells were distributed evenly throughout the lamina propria. Increased submucosal immunoreactivity was found only in Crohn's disease, in which TNF alpha positive macrophages tended to cluster around arterioles and venules, often infiltrating and disrupting vascular endothelium. It is suggested that this degree of TNF alpha production probably contributes significantly to the pathogenesis of both Crohn's disease and ulcerative colitis, by impairing the integrity of epithelial and endothelial membranes, increasing inflammatory cell recruitment, and by prothrombotic effects on the vascular endothelium.     

Coexpression of CD4 and CD8 on peripheral blood T cells and lamina propria T cells in inflammatory bowel disease by two colour immunofluorescence and flow cytometric analysis.

Using two colour immunofluorescence with fluorescein isothiocyanate and phycoerythrin labelled monoclonal antibodies and multiparameter flow cytometry, we investigated the coexpression of CD4 and CD8 antigens on peripheral blood lymphocytes and lamina propria lymphocytes of patients with ulcerative colitis and Crohn's disease and normal control subjects. Both the absolute number and the proportion of peripheral blood CD4+, CD8+ cells in inflammatory bowel disease were small but significantly increased compared with those in normal control subjects. Peripheral blood lymphocytes activated with phytohaemagglutinin showed appreciably increased coexpression of CD4+, CD8+. These CD4, CD8 positive cells were large and granular. Thus the increased number of peripheral blood CD4+, CD8+ cells in inflammatory bowel disease suggests that chronic immune activation occurs not only in the active state of the disease but also in remission. The proportion of CD4+, CD8+ cells in the lamina propria was greater than in peripheral blood in normal subjects, suggesting chronic immune stimulation of the local immune system. This was also seen in patients with Crohn's disease or inactive ulcerative colitis. The proportion of CD4+, CD8+ cells was, however, significantly less in the lamina propria of patients with active ulcerative colitis. Whether this implies a possible defect in mucosal immunoregulation in active ulcerative colitis cannot be determined from these results.