The optimized capsid gene of porcine circovirus type 2 expressed in yeast forms virus-like particles and elicits antibody responses in mice fed with recombinant yeast extracts

Porcine circovirus type 2 (PCV2)-associated diseases are considered to be the biggest problem for the worldwide swine industry. The PCV2 capsid protein (Cap) is an important antigen for development of vaccines. At present, most anti-PCV2 vaccines are produced as injectable formulations. Although effective, these vaccines have certain drawbacks, including stress with concomitant immunosuppresion, and involve laborious and time-consuming procedures. In this study, Saccharomyces cerevisiae was used as a vehicle to deliver PCV2 antigen in a preliminary attempt to develop an oral vaccine, and its immunogenic potential in mice was tested after oral gavage-mediated delivery. The cap gene with a yeast-optimized codon usage sequence (opt-cap) was chemically synthesized and cloned into Escherichia coli/Saccharomyces cerevisiae shuttle vector, pYES2, under the control of the Gal1 promoter. Intracellular expression of the Cap protein was confirmed by Western blot analysis and its antigenic properties were compared with those of baculovirus/insect cell-produced Cap protein derived from the native PCV2 cap gene. It was further demonstrated by electron micrography that the yeast-derived PCV2 Cap protein self-assembles into virus-like particles (VLPs) that are morphologically and antigenically similar to insect cell-derived VLPs. Feeding raw yeast extract containing Cap protein to mice elicited both serum- and fecal-specific antibodies against the antigen. These results show that it is feasible to use S. cerevisiae as a safe and simple system to produce PCV2 virus-like particles, and that oral yeast-mediated antigen delivery is an alternative strategy to efficiently induce anti-PCV2 antibodies in a mouse model, which is worthy of further investigation in swine.     

Combined therapeutic effect of a monoclonal anti-idiotype tumor vaccine against NeuGc-containing gangliosides with chemotherapy in a breast carcinoma model

Anti-idiotypic monoclonal antibodies (mAb) have been evaluated for actively induced immunotherapy with encouraging results. However, rational combination of cancer vaccines with chemotherapy may improve the therapeutic efficacy of these two approaches used separately. The main objective of this study was to evaluate the antitumor effect of the co-administration of 1E10 (Racotumomab), a monoclonal anti-idiotype tumor vaccine against an IgM mAb, named P3 that reacts specifically with NeuGc-containing gangliosides and low-dose Cyclophosphamide in a mammary carcinoma model. F3II tumor-bearing mice were immunized subcutaneously with 100 μg of 1E10 mAb in Alum or with 150 mg/m² of Cyclophosphamide intravenously 7 days after the tumor inoculation. While a limited antitumor effect was induced by a single 1E10 mAb immunization; its co-administration with low-dose Cyclophosphamide reduced significantly the F3II mammary carcinoma growth. That response was comparable with the co-administration of the standard high-dose chemotherapy for breast cancer based on 60 mg/m² of Doxorubicin and 600 mg/m² of Cyclophosphamide, without toxicity signs. Combinatorial chemo-immunotherapy promoted the CD8⁺ lymphocytes tumor infiltration and enhanced tumor apoptosis. Furthermore, 1E10 mAb immunization potentiated the antiangiogenic effect of low-dose Cyclophosphamide. Additionally, splenic myeloid cells Gr1⁺/CD11b⁺ associated with a suppressor phenotype were significantly reduced in F3II tumor-bearing mice immunized with 1E10 mAb alone or in combination with low-dose Cyclophosphamide. This data may provide a rational for chemo-immunotherapy combinations with potential medical implications in breast cancer.     

The Collaborative Study of Obesity and Diabetes in Adults (CODA) project: meta-analysis design and description of participating studies

The Collaborative Study of Obesity and Diabetes in Adults (CODA) project was formed to establish an international database of studies of abdominal obesity and type 2 diabetes mellitus (T2DM), and to provide analyses of these associations using individual participant data (IPD) meta-analytic techniques. The collaboration involves obtaining raw data from existing studies. The main objectives of the collaboration are to assess which simple anthropometric indices most closely predict the risk of T2DM in adults, and to investigate ethnicity and other factors that potentially modify that prediction. A second task related to primary prevention of diabetes subsequently evolved, the CODA-2 project, and is concerned with population-based methods to identify people most likely to benefit from diabetes interventions. This article describes the meta-analysis design and the studies involved. The collaboration currently has 37 studies enrolled, providing data on 260,000 participants. The proposed IPD meta-analyses will help resolve several outstanding issues in diabetes.     

Ⅰ.Nogo在神经再生中的作用

体内大部分组织如肌肉、皮肤、肝脏和外周神经,损伤后均有很强的再生能力.然而,中枢神经系统(CNS)几乎没有这种能力,损伤后的轴突及神经元不能再生.导致损伤后功能迟迟得不到恢复,如脊髓损伤导致的瘫痪.     

Acute stress and dexamethasone rapidly increase hippocampal somatostatin synthesis and release from the dentate gyrus hilus

Somatostatin is a neuropeptide whose facilitatory action in the generation of long‐term potentiation (LTP) in the hippocampal dentate gyrus has been associated with memory processes. Since stress and memory seem to share some neural pathways, we studied somatostatin release from dentate gyrus hilar cells of the hippocampus in unanesthetized free‐moving rats subjected to stress or dexamethasone treatments. In parallel, the number of dentate gyrus hilar cells expressing somatostatin mRNA was quantified by nonradioactive in situ hybridization in these two experimental conditions. Rats were stereotaxically implanted with a push‐pull cannula in the dentate gyrus hilar region. Animals were perfused 1 week later in basal or stress (30 min immobilization stress) conditions. The other group was intraperitoneally injected with the synthetic glucocorticoid dexamethasone (3 mg/kg b.w.). Samples were collected every 15 min for somatostatin radioimmunoassay. In parallel, in other groups of animals undergoing the same treatments, brains were removed for in situ hybridization studies with an oligonucleotide labeled with digoxigenin that recognizes somatostatin‐14. The results showed that stress induced a significant increase in somatostatin release from dentate gyrus hilar cells 30–45 min after immobilization stress application. Dexamethasone‐injected animals exhibited a similar response 45 min after drug administration. In situ hybridization analysis revealed that the two treatments significantly increased the number of cells expressing somatostatin mRNA in the hilar region. In conclusion, somatostatin interneurons of the hippocampal hilar region appear to be a novel stress stimulus target. Their rapid reactivity, expressed as modifications of both somatostatin release and number of cells expressing somatostatin mRNA, provides an interesting model of neuronal plasticity. Hippocampus 2001;11:469–477. © 2001 Wiley‐Liss, Inc.     

Pyogenic granuloma over the venous component of a mixed vascular malformation

A 52‐year‐old man presented with a rapidly growing red tumor on the central neckline. It had appeared over a congenital flat and pinkish vascular lesion that involved the shoulder and the upper anterior area of his chest. Intermingled with the pinkish stain, there were also some blue nodules several millimeters in diameter. Histopathologic examination revealed that the full lesion was a mixed venous‐capillary malformation. The red tumor was excised and diagnosed as a pyogenic granuloma developing over the venous component of the vascular malformation. To our knowledge, a pyogenic granuloma growing over a venous malformation has not been previously described.