牛黄解毒片的三维高效液相色谱法鉴定和指标成分的定量

用三维高效液相色谱(three dimensional HPLC)法对中成药牛黄解毒片进行了分析。结果表明该法用于牛黄解毒片中单味中药鉴定及指标成分(marker substance)含量测定均得到较满意结果。     

Anti-idiotypic antibodies specific for a pathologic anti-Pr2 cold agglutinin

The heterogeneity of human red cell (RBC) autoantibodies may be assessed by using anti-idiotypic antibodies. In this study, mouse monoclonal anti-idiotypic antibodies were produced against a pathologic RBC autoantibody with anti-Pr2 specificity. Epstein-Barr virus-transformed B-cell clones were established from a patient who had splenic lymphoma and associated immune hemolysis due to an anti-Pr2 cold autoantibody. Two of the eight clones producing this autoantibody were used to immunize mice for the establishment of hybridomas, and four monoclonal anti-idiotypic antibodies were isolated (2 IgG1 kappa and 2 IgM kappa). By the use of these anti-idiotypic antibodies, strong crossreactivity was seen on enzyme-linked immunosorbent assay with other anti-Pr2-producing clones from the same patient, but no cross-reactivity was seen with RBC autoantibodies from other individuals having anti-Pr or different specificities. Each of the anti-idiotypic antibodies inhibited hemagglutination (HA) by the patient's anti-Pr2 but failed to inhibit HA by antisera of a different RBC specificity. Cross-competition experiments indicated that all of the anti-idiotypic antibodies may recognize the same or a closely related idiotope on the anti-Pr2 autoantibody. These studies suggested that the four anti-idiotypic antibodies are directed against the same (or closely related) idiotypic determinant(s), unique to this patient's anti-Pr2 and located at or near the antigen-binding site. These anti-idiotypic antibodies may be useful tools for the study of this autoimmune response or for the development of immune therapeutic agents.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dual channel optical tomographic imaging of leukocyte recruitment and protease activity in the healing myocardial infarct

Inflammatory responses after myocardial infarction profoundly impact tissue repair. Yet, efficient tools to serially and noninvasively assess cellular and molecular functions in postinfarct inflammation are lacking. Here we use multichannel fluorescent molecular tomography (FMT) for spatiotemporal resolution of phagocytic and proteolytic activities mediated by macrophages and neutrophils in murine infarcts. We performed FMT imaging to compare the course of efficient and impaired healing in wild-type and FXIII-/- mice, respectively. Mice subjected to coronary ligation received simultaneous injections with Prosense-680, an activatable fluorescence sensor reporting on cathepsin activity, and CLIO-VT750, a magneto-fluorescent nanoparticle for imaging of phagocyte recruitment. On FMT, Prosense-680 infarct signal was 19-fold higher than background (P<0.05). Protease activity was higher in the infarcted lateral wall than in the remote, uninjured septum on ex vivo fluorescence reflectance imaging (contrast to noise ratio 118+/-24). CLIO-VT750 FMT signal coregistered with contrast enhancement in the hypokinetic infarct on MRI. Microscopic fluorescence signal colocalized with immunoreactive staining for cathepsin, macrophages and neutrophils. Flow cytometry of digested infarcts revealed monocytes/macrophages and neutrophils as the source of the fluorescence signal. Phagocytic activity peaked on day 6, and proteolytic activity peaked on day 4 after myocardial infarction. FMT detected impaired recruitment of phagocytes and protease activity in FXIII-/- mice (P<0.05). FMT is a promising noninvasive molecular imaging approach to characterize infarct healing. Spectrally resolved imaging agents allow for simultaneous assesment of key processes of in vivo cellular functions. Specifically, we show that in vivo FMT detects impaired healing in FXIII-/- mice.     

Monocyte accumulation in mouse atherogenesis is progressive and proportional to extent of disease

Monocytes participate importantly in the pathogenesis of atherosclerosis, but their spatial and temporal recruitment from circulation remains uncertain. This study tests the hypothesis that monocyte accumulation in atheroma correlates with the extent of disease by using a sensitive and simple quantitative assay that allows tracking of highly enriched populations of blood monocytes. A two-step isolation method yielded viable and functionally intact highly enriched peripheral blood monocyte populations (>90%). Recipient mice received syngeneic monocytes labeled in two ways: by transgenically expressing EGFP or with a radioactive tracer [(111)In]oxine. After 5 days, more labeled cells accumulated in the aorta, principally the aortic root and ascending aorta, of 10-wk-old ApoE(-/-) compared with 10-wk-old C57BL/6 mice (223 +/- 3 vs. 87 +/- 22 cells per aorta). Considerably more monocytes accumulated in 20-wk-old ApoE(-/-) mice on either chow (314 +/- 41 cells) or high-cholesterol diet (395 +/- 53 cells). Fifty-week-old ApoE(-/-) mice accumulated even more monocytes in the aortic root, ascending aorta, and thoracic aorta after both chow (503 +/- 67 cells) or high-cholesterol diet (648 +/- 81 cells). Labeled monocyte content in the aorta consistently correlated with lesion surface area. These data indicate that monocytes accumulate continuously during atheroma formation, accumulation increases in proportion to lesion size, and recruitment is augmented with hypercholesterolemia. These results provide insights into mechanisms of atherogenesis and have implications for the duration of therapies directed at leukocyte recruitment.     

Inhalation of a harmless antigen (ovalbumin) elicits immune activation but divergent immunoglobulin and cytokine activities in mice

BACKGROUND: Exposure to aerosolized harmless antigen such as ovalbumin (OVA) has previously been shown to induce inhalation tolerance, a state characterized by inhibition of IgE synthesis and airway inflammation, upon secondary immunogenic antigen encounter. Immune events associated with this phenomenon are still poorly understood. OBJECTIVE: The aim of this study was to investigate cellular and molecular mechanisms underlying this state of 'unresponsiveness'. METHODS: After initial repeated OVA exposure, mice were subjected to a protocol of antigen-induced airway inflammation, encompassing two intraperitoneal injections of OVA adsorbed to aluminium hydroxide followed by airway challenge. We assessed immune events in the draining lymph nodes after sensitization, and in the lungs after challenge. RESULTS: In animals initially exposed to OVA, we observed, at the time of sensitization, considerable expansion of T cells, many of which expressed the activation markers CD69 and CD25, as well as increased numbers of antigen-presenting cells, particularly B cells. While these animals produced low levels of IgE, the observed elevated levels of IgG1 signified isotype switching. Splenocytes and lymph node cells from OVA-exposed mice produced low levels of IL-4, IL-5, IL-13 and IFN-gamma, indicating aborted effector function of both T helper (Th)2- and Th1-associated cytokines. Real time quantitative polymerase chain reaction (PCR) (TaqMan) analysis of costimulatory molecules in the lungs after in vivo challenge showed that B7.1, B7.2, CD28 and CTLA-4 mRNA expression was low in animals initially exposed to OVA. Ultimately, these events were associated with abrogated airway inflammation and attenuated airway hyper-responsiveness. The decreased inflammation was antigen-specific and independent of IL-10 or IFN-gamma. CONCLUSION: Initial exposure to OVA establishes a programme that prevents the generation of intact, fully functional inflammatory responses upon secondary antigen encounter. The absence of inflammation, however, is not associated with categorical immune unresponsiveness.     

Early clinical manifestation of glutaric aciduria type I and nephrotic syndrome during the first months of life

We describe a male patient with glutaric aciduria type I which had already presented during the neonatal period with therapy-resistant seizures. In the course of the disease, he also developed choreoathetosis and dystonia. The disease was associated with nephrotic syndrome. Renal histology showed signs of a glomerular disorder with shrinking of glomerular tufts, increase in mesangial matrix, proliferation of extracapillary epithelial cells and formation of larger epithelial crescents. The child died at 22 weeks of age due to end-stage renal failure. This report illustrates an unusual and early clinical manifestation of glutaric aciduria type I and a hitherto unknown association with nephrotic syndrome in early childhood.