Transferrin receptor distribution and iron deposition in the hepatic fobufe of iron-overloaded rats

Under the condition of obvious iron-overload, there is a zonal hernoeiderin (iron) deposition in hepatic lobules. The deposition is heavtest in the periporfal (zone 1) and lightest in the perivenws (zone 3) hepatocytes. However, the mechanism for this pattern of iron deposition is obscure. Hepatic tissues from control, iron-deficlent or ironoverloaded Wistar rats me used to study its pathogenesis. iron-deficiency was Induced by a low Iron regimen. Ironoverload was produced by repeated intraperitoneal injections of ferric nitrilotriace-We (Fe³⁺-NTA) for 1–4 months. Liver tissues of the rats were lmmunohistochemically and histochemically stained for tmnaferrin receptor (TfR), transferrin (Tf), ferritin (Ft), and iron. The staining intensity of TfR, Tf and Ft increased in hepatocytes of iron-deflctent rats and decreased in that of the iromverloaded in comparison with the control rats. TfR atalning was strong in zone 1, with gradual transition into weak staining in zone 3 hepatocytes of the rat liver. TfR located primarily on the hepatocyte membrane. Tf had both membranous and cytoplasmic distribution. Many hepatocytes in group B had strong cytoplasmic Tf staining. Conversely, only a few hepatocytes had weakly stained cytoplasmic Tf in group C. Hepatocytes and Kupffer cells were Ft positive in control rats. Ft was distributed only in the cytoplasm. The staining intensity of Ft was stronger in zone 3 than in zone 1 hepatocytes of iron-deficient rats. In iron-overloaded rats, the iron deposition was severe in zone 1 and mild in zone 3 hepatocytes. These findings suggest that uptake of iron into hepatocytes in vivo is regulated and mediated by TfR and Tf. Gradient TfR distribution from zone 1 to 3 hepatocytes and active TfR-Tf mediated iron uptake resuited in the zonal iron deposition in the hepatic lobule of iron-overloaded rats.     

Health status, adherence with health recommendations, self-efficacy and social support in patients with rheumatoid arthritis

A study was performed in 86 patients with rheumatoid arthritis (RA) to assess their health problems, the problems they experience in adhering to health recommendations and the relationships of these problems with self-efficacy and social support. Feeling dependent, disability and pain were the most important health related problems. The results showed self-efficacy to be related to the subjective experience of health status as measured by DUTCH-AIMS. Social emotional support was not related to health status and contrary to what we expected social instrumental support was positively related to health status. The majority of the patients (55%) experienced adherence problems with health recommendations. These problems were not related to functional incapacity, pain or other aspects of health status but to the patient's self-efficacy expectations about coping with arthritis. Our conclusion is that to improve the self-management of disability and pain and adherence to health recommendations, patient education should be aimed at strengthening self-efficacy expectations in which social emotional support might be a motivating factor.     

Respiratory mechanics and pleural remodelling in pleurodesis induced by barium sulphate

The aim of this study was to determine whether an intrapleural injection of barium sulphate would produce pleurodesis in rats. Additionally, respiratory mechanics and pleural remodelling were analysed. Single intrapleural injection of barium sulphate (100%) or saline was given to Wistar rats. Respiratory system, lung, and chest wall elastic, resistive and viscoelastic/inhomogeneous pressures were measured by the end-inflation occlusion method at 2 and 30 days after injection. The pleura were examined for gross and histopathological evidence of pleural inflammation and fibrosis, and the underlying lungs were also studied by morphometry. All pulmonary mechanical parameters increased at day 2, but were not different from control at 30 days after injection. Chest wall mechanical parameters did not change. Macroscopic evaluation demonstrated pleural adherence without haemothorax. Histopathologic analysis showed pleural inflammation and fibrosis. There was no alveolar inflammation or fibrosis in both groups. In conclusion, barium sulphate induced pleurodesis with either no changes in respiratory mechanics or lung lesion at day 30.     

Quantitation of aberrant interlocus T-cell receptor rearrangements in mouse thymocytes and the effect of the herbicide 2,4-dichlorophenoxyacetic acid

Small studies in human populations have suggested a correlation between the frequency of errors in antigen receptor gene assembly and lymphoid malignancy risk. In particular, agricultural workers exposed to pesticides have both an increased risk for lymphoma and an increased frequency of errors in antigen receptor gene assembly. In order to further investigate the potential of such errors to serve as a mechanistically based biomarker of lymphoid cancer risk, we have developed a sensitive PCR assay for quantifying errors of V(D)J recombination in the thymocytes of mice. This assay measures interlocus rearrangements between two T-cell receptor loci, V-gamma and J-beta, located on chromosomes 13 and 6, respectively. The baseline frequency in four strains of mice was determined at several ages (2-8 weeks of age) and was found to be stable at approximately 1.5 x 10(-5) per thymocyte. Strain AKR, which has a high susceptibility to T-cell lymphomas, did not show an elevated frequency of aberrant V(D)J events. We used this assay to examine the effects of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) on the frequency of these events. Female B6C3F1 mice, 27 days of age, were exposed to 2,4-D by gavage at doses of 0, 3, 10, 30, and 100 mg/kg/day for 4 successive days and sacrificed on day 5. Thymus DNA was isolated and examined for illegitimate V(D)J recombination-mediated gene rearrangements. In addition, pregnant mice were exposed to 2,4-D and thymocytes from the offspring examined at 2 weeks of age. No significant increase in aberrant V(D)J rearrangements was found, indicating that under these conditions 2,4-D does not appear to effect this important mechanism of carcinogenesis.     

Activation of invariant NK T cells protects against experimental rheumatoid arthritis by an IL-10-dependent pathway

Invariant natural killer T (iNKT) cells are a unique lymphocyte subtype implicated in the regulation of autoimmunity and a good source of protective Th2 cytokines. Agonist alpha-galactosylceramide (alpha-GalCer) of iNKT cells exert a therapeutical effect in type 1 diabetes. We investigated whether iNKT activation with alpha-GalCer was protective in collagen-induced arthritis (CIA) in DBA/1 mice, a standard model of rheumatoid arthritis. Here, we have shown that in vivo iNKT cell function was altered in DBA/1 mice since stimulation with alpha-GalCer led to decreased IL-4 and IFN-gamma levels in sera, as compared with C57BL/6 mice. alpha-GalCer induced a clear-cut diminution of clinical and histological arthritides. An anti-IL-10 receptor antibody abrogated the protective effect of alpha-GalCer, suggesting a key role for IL-10 in the protection against CIA by activated iNKT cells. Confirming these data, disease protection conferred by alpha-GalCer correlated with the ability of LN CD4+ cells to secrete larger amounts of IL-10. These findings suggest that in CIA susceptibility to autoimmunity is associated with dysfunctions of iNKT cells. Our demonstration that iNKT cell activation by alpha-GalCer remains efficient in CIA-prone DBA/1 mice to provide protective IL-10 suggests that this could be used therapeutically to treat autoimmune arthritis.     

Regulation of CCR5 expression and MIP-1alpha production in CD4+ T cells from patients with rheumatoid arthritis

Production of CCR5 expression and MIP-1alpha, a ligand of CCR5, by CD4+ T cells from patients with rheumatoid arthritis (RA) were studied. We analysed further the influence of IL-15 stimulation, CD40/CD40 ligand (CD40L) interaction and CCR5 promotor polymorphism. One hundred and fifty-five RA patients and another 155 age- and sex-matched healthy individuals were enrolled. Peripheral CD4+ and double negative (DN) T cells from patients had lower portions of CCR5, whereas synovial CD4+ and DN T cells showed a much higher CCR5 expression. IL-15 significantly up-regulated the expression of CCR5 on purified CD4+ T cells. CD40L expression on synovial CD4+ T cells was increased greatly in CCR5+ portions by IL-15. MIP-1alpha production by synovial CD4+ T cells was also enhanced by IL-15. Co-culture of CD40 expressing synovial fibroblasts with IL-15-activated synovial CD4+ T cells significantly increased MIP-1alpha production. Expression of CCR5 on patients' CD4+ T cells was not influenced by the promotor polymorphism of CCR5 gene. Taken together, these data suggest CCR5+CD4+ T cells infiltrate the inflamed synovium and IL-15 up-regulates CCR5 and CD40L expression further and enhance MIP-1alpha production in synovial CD4+ T cells. Production of MIP-1alpha by synovial fibroblasts is significantly increased by engagement of CD40 with CD40L. Synovial microenvironment plays a potential role in regulation of CCR5+CD4+ T cells in rheumatoid joints.     

Analysis of fMRI data by blind separation of data in a tiny spatial domain into independent temporal component

Independent Component Analysis (ICA) is a promising tool for the analysis of functional magnetic resonance imaging (fMRI) time series. In these studies, mostly assumed is a spatially independent component map of fMRI data (spatial ICA). In this paper, we assume that the temporal courses of the signal and noises are independent within a Tiny spatial domain (temporal ICA). Then with fast-ICA algorithm, spatially neighboring fMRI data were blindly separated into several temporal courses and were preassumed to be formed by a signal time course and several noise time courses where the signal has the largest correlation coefficient with the reference signal. The final functional imaging was completed for the signals obtained from each voxel. Simulations showed that compared with the spatial ICA method, the new temporal ICA method is more effective than the spatial ICA in detecting weak signal in a fMRI dataset. As background noise, the simulations include simulated Gaussian noise and fMRI data without stimulation. Finally, vivo fMRI tests showed that the excited areas evoked by a visual stimuli are mainly in the region of the primary visual cortex and that evoked by auditory stimuli are mainly in the region of the primary temporal cortex.     

大鼠舌咽和迷走神经感觉神经节内P2X_2、P2X_3受体与calretinin和IB4结合位点的共存

应用免疫组织化学荧光三标方法结合激光共聚焦显微镜技术研究了大鼠舌咽和神经迷走神经感觉神经节内ATP受体P2X2、P2X3与calretinin、植物凝集素(IB4)结合位点的共存。结果显示:舌咽和迷走神经感觉神经节内可见大量P2X2、P2X3阳性的神经元胞体和纤维,P2X2受体阳性细胞多为大、中、小型神经元,P2X3阳性细胞多为中、小型神经元;可见较多的P2X2/IB4及P2X3/IB4双标神经元。有(91.1±4.7)%(结状神经节)、(78.8±2.4)%(岩神经节)、(76.8±2.7)%(颈静脉神经节)为P2X2阳性细胞;(77.0±3.2)%(结状神经节)、(91.2±3.9)%(岩神经节)、(78.4±3.6)%(颈静脉神经节)的P2X3阳性细胞结合IB4;有8.9±1.6%(结状神经节)、7.7±1.4%(岩神经节)、9.1±1.1%(颈静脉神经节)的P2X2阳性细胞呈calretinin阳性;三神经节内均观察到少量P2X2/calretinin/IB4三标细胞。P2X3/calretinin双标或P2X3/calretinin/IB4三标神经元数量较少。这些结果提示,舌咽和迷走神经感觉神经节内ATP受体P2X2、P2X3与IB4结合位点存在广泛的共存,部分P2X2受体与calretinin存在着共存。     

β3‐Adrenoceptors modulate left ventricular relaxation in the rat heart via the NO‐cGMP‐PKG pathway

Aims: Using a model of isolated and Langendorff‐perfused rat heart we analysed whether activation of β₃‐adrenergic receptors (β₃‐ARs) influences ventricular lusitropic performance. We also focused on the NOS/NO/cGMP/PKG cascade as the signal transduction mechanism. Methods: Hearts were treated with increasing concentrations (from 10^?12 to 10^?6 m ) of BRL₃₇₃₄₄, a selective β₃‐AR agonist, and cardiac performance was evaluated by analysing both lusitropic parameters and coronary motility. Cardiac preparations were also perfused with BRL₃₇₃₄₄ in the presence of either isoproterenol (ISO) or nadolol, or pertussis toxin (PTx), or selective inhibitors of the NOS/NO/cGMP/PKG pathway. Results: BRL₃₇₃₄₄ caused a significant concentration‐dependent reduction in (LVdP/dt)_min, a decrease in half time relaxation significant starting from 10^?12 m , and an increase in (LVdP/dt)_max/(LVdP/dt)_min ratio (T/?t). BRL₃₇₃₄₄ abolished the ISO‐mediated positive lusitropism. β₃‐AR‐dependent effects on relaxation were insensitive to β₁/β₂‐AR inhibition by nadolol (100 nm ), and were abolished by G_i/o protein inhibition by PTx (0.01 nm ). NO scavenging by haemoglobin (10 μm ), and nitric oxide synthase (NOS) inhibition by NG‐monomethyl‐l ‐arginine (10 μm ) revealed the involvement of NO signalling in BRL₃₇₃₄₄ response. Pre‐treatment with inhibitors of either soluble guanylate cyclase (ODQ; 10 μm ) or PKG (KT₅₈₂₃; 100 nm ) abolished β₃‐AR‐dependent negative lusitropism. In contrast, anantin (10 nm ), an inhibitor of particulate guanylate cyclase, did not modify the effect of BRL₃₇₃₄₄ on relaxation. Conclusion: Taken together, our findings provide functional evidence for β₃‐AR modulation of ventricular relaxation in the rat heart which involves PTx‐sensitive inhibitory Gi protein and occurs via an NO‐cGMP‐PKG cascade. Whether the effects of β₃‐AR stimulation on lusitropism are beneficial or detrimental remains to be established.     

大鼠视前内侧区的传出性神经纤维联系—WGA—HRP法研究

应用WGA-HRP顺行轴突运输研究大鼠视前内侧区传出性神经纤维投射。结果表明:视前内侧区的上行投射向嘴侧经斜角带进入外侧隔核;经髓纹进入缰核;经无名质进入杏仁前区及经终纹进入杏仁内侧核,另有标记纤维经内侧前脑束向外下行,经视束上方进入杏仁内侧核。下行投射经内侧前脑束进入下丘脑室旁核、外侧区、内侧核、后核、弓状核、乳头体前腹核和乳头体上核。继续向尾侧,标记纤维进入中脑腹侧背盖区,并投射到中缝正中核及中缝背核。