T-cell specificity in murine autoimmune haemolytic anaemia induced by rat red blood cells

Autoimmune haemolytic anaemia (AIHA) can be induced in mice by repeated injections with rat red blood cells (RBC). Here we describe the identification of rat and murine RBC antigens recognized by T-cells from mice with this disease. Splenic T-cells from mice with AIHA proliferated in response to multiple murine RBC membrane components, each of which is recognized by rat RBC induced autoantibodies. Thus, there were responses to murine autoantigen fractions that correspond in apparent molecular mass with the anion channel Band 3, with spectrin from the membrane skeleton and with the high and low molecular mass glycophorins, and the equivalent fractions from rat RBC also stimulated proliferation by T-cells. It was confirmed that purified Band 3 from murine and rat RBC also elicited responses. In contrast with the results in AIHA, T-cells from healthy control mice failed to respond to the antigens from either species, with the exception of proliferation induced by murine spectrin in one experiment and weak responses elicited by rat Band 3. It is suggested that T-cells activated by multiple cross-reactions between rat and murine RBC proteins, and by epitope spreading, are necessary to drive autoantibody production in this model of AIHA.     

胶原基真皮再生支架的微结构控制

综述了影响胶原基真皮再生支架微结构的各种因素及其控制方法。胶原基真皮支架的生物活性和修复创面的能力受到诸如除胶原外的其它主要材料 (第二组分 )、支架的孔径和孔隙率、支架厚度、生物活性因子以及交联度等多方面因素的综合影响。从材料学、生物学和医学的角度综合地应用物理、化学和生物学的手段探索各种影响因素的控制方法是组织工程皮肤研究的重点。     

Inhibitory effect of ketamine on phosphorylation of the extracellular signal-regulated kinase1/2 following brain ischemia and reperfusion in rats with hyperglycemia

To determine if the inhibitory effects of ketamine on the extracellular signal-regulated kinase (ERK) 1/2 are involved in reduction of the hyperglycemia-exaggerated cerebral ischemic lesion, rats with normoglycemia, hyperglycemia, or hyperglycemia supplemented with ketamine were subjected to 15 min of forebrain ischemia, and then, reperfusion for 0.5, 1, and 3h. Phosphorylation of ERK1/2 in the brain tissues was assessed by immunohistochemistry and Western blot analysis. In rats with normoglycemia, we demonstrated a moderate increase of the ERK1/2 phosphorylation in the cingulum cortex and hippocampus CA3 following an ischemic intervention. It quickly dropped to control levels after reperfusion for 0.5h. In rats with hyperglycemia, however, the increase of the ERK1/2 phosphorylation in these areas was significantly higher in all animals reperfused. The neuronal death, detected by the TdT-mediated-dUTP nick end labeling assays, was found in the cingulum cortex (5.23+/-2.34, per high power feild) and hippocampus CA3 areas (6.29+/-3.68, per 1mm(2)) in hyperglycemic group after reperfusion for 3h. With ketamine treatment, the ERK1/2 phosphorylation in cingulum cortex and hippocampus CA1 and CA3 areas was found to be the same as that in normoglycemia rats. Our results suggest that hyperglycemia may increase the ischemic insult through modulation of the signal transduction pathways involving ERK1/2. The inhibitory effects of ketamine on the hyperglycemia-activated ERK1/2 phosphorylation are probably through inhibition of the N-methyl d-aspartate-mediated calcium influx, which subsequently reduce the hyperglycemia-exaggerated cerebral damage.     

Translocation of Enterococcus faecalis strains across a monolayer of polarized human enterocyte-like T84 cells

We used a two-chamber system to study transcytosis of Enterococcus faecalis across monolayers of human colon carcinoma-derived T84 cells, which show structural resemblance to the native intestine. Among 16 E. faecalis isolates from different sources, the well-characterized strain OG1RF and 8 other isolates (2 endocarditis isolates, 1 urine isolate, and all 5 fecal isolates) showed translocation in this assay, while 6 clinical isolates (3 endocarditis and 3 urine isolates), the recipient strain JH2-2, and the control, Escherichia coli DH5alpha, had no detectable translocation. Of two OG1RF mutants involving the previously studied epa (enterococcal polysaccharide antigen) gene cluster, known to be needed for virulence and resistance to killing by polymorphonuclear leukocytes, one epa mutant (TX5179) was unable to translocate, while TX5180, with an epa disruption farther downstream, showed a moderate decrease in translocation relative to that of the wild-type strain OG1RF (P < 0.01), indicating that the epa gene cluster is important for translocation across a T84 monolayer. This observation was confirmed by complementation of the epa mutant (TX5179) with epa genes and restoration of its translocation ability. In conclusion, we have demonstrated translocation of at least some strains of E. faecalis across T84 monolayers, although strains differ considerably in this ability, and we have demonstrated that epa mutations can cause marked changes in successful translocation. These results suggest that this model may be a useful in vitro system for studying the process of translocation from the intestinal tract.     

动脉粥样硬化斑块中内皮素生成及分布的免疫组织化学观察

应用免疫组化方法对人动脉粥样硬化（ＡＳ）斑块中内皮素（ＥＴ）进行分析，发现除内皮细胞外，增生的平滑肌细胞（ＳＭＣ）中也含有大量的内皮素；在内皮剥脱的大鼠胸主动脉，增生的内膜ＳＭＣ能产生丰富的内皮素。内皮素放射免疫测定证实ＳＭＣ增生的活跃程度与内皮素量呈正比。提示内皮素合成增多与ＡＳ斑块内ＳＭＣ增生关系密切。     

Molecular orientation of ultrahigh molecular weight polyethylene induced by various sliding motions

Wear and wear debris of ultrahigh molecular weight polyethylene (UHMWPE) in joint replacements have been recognized as one of the major contributors to the failure of orthopedic implants. The detailed wear mechanism of polyethylene under biomechanic motions is not well understood. In simulation wear bench tests, it was found that unidirectional sliding produces the least amount of wear, reciprocating motion increases wear significantly, and cross-shear motion (similar to hip and knee joint motion in the human body) produces the highest amount of wear. Conventional wear theories are inadequate to explain this observation. This study utilizes resonant absorption of linearly polarized soft X-rays at a synchrotron radiation beam line to measure the molecular orientation of a UHMWPE surface layer subjected to different wear motions. Carbon-K-edge partial-electron-yield X-ray absorption measurements were done on the worn UHMWPE samples. X-ray absorption measurements show conclusively that the molecular chains of UHMWPE align preferentially parallel to the direction of sliding. Examination under various wear motions showed that unidirectional shear produced the maximum chain orientation, whereas cross-shear wear motions produced the least amount of orientation. When polymeric chains align, the surface layer tends to be more brittle and hard, thus resisting wear. When they do not align, loose chains may be subjected to both Mode I and Mode II fracture, hence increasing the wear rate. This molecular alignment observation may offer an explanation of why different wear motions have different wear characteristics.     

Characterization of chenodeoxycholic acid as an endogenous antagonist of the G-coupled formyl peptide receptors

OBJECTIVE AND DESIGN: To demonstrate the role of bile acids in immune modulation we examined the ability of select bile acids to inhibit leukocyte migration and chemoattractant receptor function. MATERIALS: To elucidate this mechanism, we employed primary human monocytes, neutrophils and cell lines transfected to express either the high affinity fMLP receptor (FPR) or the low affinity fMLP receptor like 1 (FPRL1). Treatment: Cells were treated with chenodeoxycholic acid (CDCA) and related bile acids in a 0-400 micromolar range. METHOD: Cell viability, chemotaxis and calcium flux analysis were preformed. RESULTS: We observed that pathophysiological levels (< or = 150 micromolar) of CDCA competitively inhibited 3H-fMLP binding to human monocytes, FPR and FPRL1 transfected cells. Additionally, CDCA reduced both the chemotactic and calcium flux responses induced by fMLP or "W" peptide. Further, CDCA inhibited anti-FPR antibody binding to monocytes. CONCLUSIONS: CDCA selectively inhibited human leukocyte chemotaxis and calcium flux induced by fMLP, but not other chemoattractants, suggesting a mechanism for inhibition of inflammation and suppression of innate immune response.     

下颌骨的应力迹线与骨小梁构筑的关系

根据下頜光弹的等倾线图,描绘出主应力迹线规迹。在下颌侧位X线照片和去除唇侧及颊侧密质骨板的标本上,观察骨密质和松质的配布,以及骨小梁的排列和方向,试行探讨骨小梁方向与主应力迹线的关系。用光弹法求得的主应力迹线有两个系列:S_1系在下颌体近水平方向,在下颌支近垂直方向分布;S_2系力线与S_1系诸力线呈正交。在标本和X线照片上,主应力迹线规迹不同程度地在骨小梁的排列上有所反映。