Influence of insulin on cholesterol removal from macrophages and cholesterol ester uptake by HepG2 cells

The fact that an increased blood insulin level is observed in patients with coronary artery disease (CAD) confirms the hypothesis that insulin promotes the development of atherosclerosis. The low high-density lipoprotein (HDL) concentration observed in such patients may contribute to alteration in reverse cholesterol transport and promote the accumulation of sterols in vascular tissue. We examined the effect of insulin (20−1000μUmL⁻¹) on cholesterol efflux into HDL₃ particles from human blood monocyte/macrophages and rat peritoneal macrophages preloaded with labelled cholesterol esters, and the influence of insulin on the accumulation of sterols by rat liver cells and HepG2 cell line in vitro models. Insulin at concentrations up to 250μUmL⁻¹ inhibited the efflux of cholesterol from rat macrophages and promoted high uptake of sterols by both types of hepatic cells. Pharmacological concentrations higher than 250μU mL⁻¹ exerted the opposite effect. In the case of human macrophages, an insulin concentration of 20μUmL⁻¹ increased cholesterol removal, whereas 100−200μU mL⁻¹ insulin inhibited cholesterol removal from cells, and very high concentrations (>350μUmL⁻¹) again increased cholesterol removal. We have shown that insulin excess counteracts the beneficial effects of HDL in removing cellular cholesterol and, therefore, may promote development of atherogenesis.     

奥沙利铂持续腹腔热灌注治疗癌性腹水的临床研究

目的研究奥沙利铂(L-OHP)持续腹腔热灌注治疗癌性腹水的疗效和毒副作用。方法将76例癌性腹水患者随机分为三组:奥沙利铂持续腹腔热灌注化疗组(热化组)26例,引出腹水后,在加热的5%葡萄糖溶液2500～3500ml中加入L-OHP200mg/m2,持续体外循环腹腔热灌注,腹腔内温度保持在41～43℃,持续60min;腹腔内注射奥沙利铂化疗组(单化组)26例,常规腹腔穿刺引流腹水后注射L-OHP200mg/m2。单纯腹腔热灌注组(单热组)24例,加热5%葡萄糖溶液2500～3500ml,持续热灌注60min。结果总有效率(CR PR)54%(41/76)。热化组、单化组和单热组的有效率分别为76.9%(20/26)、50.0%(13/26)、33.3%(8/24),P<0.05。急性腹痛:热化组53.8%(14/26),单热组16.7%(4/24)。单化组无急性腹痛病例,P<0.05。麻痹性肠梗阻:热化组19.2%(5/26),单化组7.7%(2/26),单热组16.7%(4/24),P>0.05。结论奥沙利铂持续腹腔热灌注治疗恶性腹水是一种新的有效的治疗方法,毒副作用不大。     

OK-432增强人腹腔抗癌免疫功能的研究

目的　探讨腹腔内注射OK - 4 32增强腹腔免疫功能的机制。方法　选择非炎症和非肿瘤手术患者作为实验对象 ,实验组分别于手术前 72小时、4 8小时和 2 4小时腹腔内注射 4KE的OK - 4 32。开腹后采集腹腔内巨噬细胞 ,并用人胃癌MKN1细胞作为靶细胞对巨噬细胞的癌细胞毒性进行分析。同时采集大网膜 ,对大网膜乳斑的数量和面积进行了观察分析。结果　OK - 4 32显著增加了腹腔内巨噬细胞的数量 (P <0 .0 5 )、增强了巨噬细胞的吞噬活性和酶活性 (P <0 .0 5 )、增加了NO的分泌和巨噬细胞的癌细胞毒性 (P <0 .0 5 ) ,以及大网膜乳斑的数量和面积 (P <0 .0 5 )。结论　手术前腹腔内注射OK - 4 32可以作为预防癌细胞腹腔内种植转移的有效方法。     

The Cellular Lesion of Humoral Rejection: Predominant Recruitment of Monocytes to Peritubular and Glomerular Capillaries

Accumulation of inflammatory cells within capillaries is a common morphologic feature of humoral renal allograft rejection and is most easily appreciated if it occurs in glomeruli. The aim of our study was to determine the amount and composition of immune cells within glomeruli and peritubular capillaries (PTC) in cellular and humoral allograft rejection. Immunofluorescent double-labeling for CD31 and CD3 or CD68 was used for phenotyping and enumerating immune cells within glomeruli and PTC. The major findings are: (1) accumulation of immune cells in PTC is far more common than it would be anticipated based on the assessment by conventional histology; (2) it is not the absolute number of immune cells accumulating within capillaries, but rather the composition of the intracapillary cell population that distinguishes humoral rejection from cellular rejection and (3) in C4d positive biopsies a predominantly monocytic cell population accumulates not only within glomeruli but also within PTC. The median value of monocyte/T-cell ratio within PTC was 2.3 in C4d positive biopsies but only 1 (p = 0.0008) in C4d negative biopsies. Given their prominent presence within capillaries and their extensive biological versatility monocytes might contribute to the capillary damage observed in acute and chronic allograft rejection.     

哮喘患者CD8+T细胞对单核/巨噬细胞抗原递呈功能的影响

目的：探讨支气管哮喘患者CD8⁺T细胞对单核/巨噬细胞抗原递呈功能的影响。方法：哮喘患者20例，健康对照22例，分别取静脉血5 mL，并分离MΦ、CD8⁺T细胞和B细胞。每份血样分成4组：MΦ递呈抗原组、CD8⁺T细胞参与MΦ递呈抗原组、CD8⁺T细胞体外活化后对MΦ递呈抗原影响组及自然状态下CD8⁺T细胞对MΦ细胞递呈抗原影响组。各组用CTLL₂P抗原刺激18 h后，洗去刺激原，与自体B细胞共同孵育10 d，吸取上清液，测定特异性IgM、IgE、IgG含量。 结果:①哮喘患者MΦ单独递呈抗原，自体B细胞特异性IgM(A490值)（0.034±0.022）明显低于健康人(0.116±0.080)（P<0.05）；CD8⁺T细胞与MΦ共同培养递呈抗原时，产生的特异性IgM(A490值)(0.031±0.021)低于健康人(0.079±0.064)（P<0.05）；②哮喘患者CD8⁺T细胞与MΦ共同培养递呈抗原时，产生的特异性IgG(A490值)(0.102±0.041)明显高于健康人(0.081±0.067)（P<0.05），自然状态下及体外活化后CD8⁺T细胞与递呈抗原MΦ共同培养，产生的特异性IgG(A490值)(0.105±0.066, 0.079±0.059)与健康人(0.066±0.038, 0.069±0.047)无明显差别(P>0.05)；③哮喘患者CD8⁺T细胞与MΦ共同培养递呈抗原产生的特异性IgE(A490值)(0.171±0.154)高于健康人(0.147±0.059)（P<0.05）。 结论:哮喘患者CD8⁺T细胞对MΦ递呈抗原产生免疫球蛋白有调节作用,而且参与哮喘发病。     

Sodium removal and sodium concentration during peritoneal dialysis: effects of three methods of sodium measurement.

BACKGROUND: Sodium removal (NaR) may have a major impact on the survival of peritoneal dialysis patients. The dialysate/plasma sodium concentration ratio (D/P(Na)) is an indirect index of transcellular water transport by aquaporin channels, and thus of ultrafiltration. Sodium concentration can be assessed by means of flame photometry (F), and direct (D-ISE) or indirect ion-selective electrodes (I-ISE), but these methods have different properties. I-ISE is being used increasingly in clinical laboratories. The aim of this study was to evaluate NaR and D/P(Na) using the three different measurement methods. METHODS: We performed peritoneal equilibration tests (PETs) in 44 peritoneal dialysis patients and calculated the NaR. We also calculated D/P(Na) during the test; plasma and dialysate sodium concentrations were measured by F, D-ISE and I-ISE. RESULTS: NaR was lower (P<0.001) with D-ISE (69+/-29 mmol) than with F (81+/-29 mmol) or I-ISE (79+/-28 mmol). D/P(Na) was also lower at baseline (0.92+/-0.02 vs 0.95+/-0.02 and 0.95+/-0.02; P<0.001), after 60 min (0.87+/-0.03 vs 0.90+/-0.03 and 0.90+/-0.03; P<0.001) and at the end of PET (0.88+/-0.04 vs 0.92+/-0.04 and 0.92+/-0.04; P<0.001) when measured by D-ISE in comparison with F and I-ISE, respectively. CONCLUSIONS: NaR and D/P(Na) were lower when measured by the D-ISE method compared with the F and I-ISE methods. NaR and D/P(Na) were similar when measured by F or I-ISE. I-ISE can be used reliably in the evaluation of NaR and D/P(Na) in everyday clinical practice of peritoneal dialysis.     

Macrophage response to microtextured silicone

Seven different silicone surface textures were tested for effect on macrophage spreading and metabolic activity in vitro. Variables of the textured arrays that could modify spreading were determined to be the size, spacing between, depth, density, and orientation of the individual surface events and the roughness of the surfaces. Cells were influenced by the size of the events and the roughness of the surfaces more than any other variables. Cell morphology data, surface area and perimeter, could be divided into discrete regions that correlated well with the size of the events. Cell dimensions on 5μm textures were smallest while those on smooth silicone and glass surfaces were the largest. Surface texture events may be modifying contact guidance of the cells or interacting with specific transmembrane proteins to alter cell shape and function. The mitochondrial activity of cells attached to the textured silicones was determined by measuring the amount of reduced MTT directly through live cells. Cells on polystyrene (PS), 5VP and 8VP textures were metabolically more active than cells on the other textures. PMA was used to stimulate cells on the various textures. PMA-stimulated cells, on the smaller textures, 2VP, 5VP and 5CP, were less active than test cells that were not stimulated. The inability of PMA to stimulate these cells may be due to a structural alteration of protein kinase C. An hypothesis is introduced that includes a possible mechanism of how a micrometre-sized surface texture could modify cell function.     

Immunopathology of ocular sarcoidosis

Summary Sarcoidosis is a multisystem disease characterized by enhanced immune responses at sites of involvement. To elucidate the immunopathogenesis of ophthalmic lesions, cell infiltrates in biopsies from conjunctiva and other tissues involved (lungs, lymph nodes, skin) were studied in 26 patients with active sarcoidosis in order to define the surface phenotype and the distribution of cells in granulomatous lesions. Biopsy specimens were also stained for detection of immunoglobulins, complement and fibrinogen deposits. The data demonstrate a lymphocytes/macrophages interaction in the central core of granulomatous areas as the crucial event that initiates the maintains the state of inflammation: at all sites of disease activity is present a compartmentalization of T-cells expressing a helper-related phenotype which account for the great majority of infiltrating cells both in the early lesions (aggregate of macrophages surrounded by lymphocytic infiltrate) and in well-organized sarcoid granulomata. The presence of plasma cells and immunoglobulin deposits may represent an epiphenomenon in line with the helper infiltration, suggesting a local hyper-reactivity of the B-cells immune system. This study suggests some immunopathogenetic mechanisms leading to the formation and growth of conjunctival sarcoid granulomata.     

Adherence of Staphylococcus aureus to cultures of human peritoneal mesothelial cells

Strains of Staphylococcus aureus, isolated from the effluentof patients with peritonitis on CAPD (continuous ambulatoryperitoneal dialysis), adhered well to both cultured human mesothelialcells and to fibronectin, but not to laminin or gelatin. Mesothelialcells grown in medium M199 exhibited more surface fibronectincompared to cells grown in MEM-Dval and demonstrated higherlevels of S. aureus adherence. Soluble fibronectin concentrations up to lOµg/ml increasedthe adherence of S. aureusto cultured mesothelial cells. Thedose-response curve was consistent with the binding of fibronectinto a saturable receptor of apparent dissociation constant (K_D)= 1.7xlO^–10 M. This corresponds closely to the K_D (2xlO^–10M) of the staphylococcal fibronectin-binding protein. S. aureus adherence was increased following the preincubationof mesothelial cell monolayers with interleukin-1 and was maximalafter 6 h preincubation. Treating mesothelial cells with interferon-gammafor 48–72 h reduced the adherence of S. aureus.