Partial purification and osteoinductive activity of swine bone morphogenetic protein.

Bone morphogenetic protein (BMP) was isolated from the bone matrix of swine and partially purified by means of differential precipitation and molecular sieve chromatography. The molecular weight of BMP estimated by SDS-gel electrophoresis was 19,000 dalton. Bioassay by implanting two milligrams of BMP fraction into thigh muscles of mice resulted in bone formation in 100% of the experimental animals.

     

Reduced expression of CD69 and adhesion molecules of T lymphocytes in asthmatic children receiving immunotherapy

T lymphocytes play a fundamental role in the initiation and regulation of chronic inflammatory responses in patients with asthma. CD69 is an early marker of T‐cell activation. The levels of intercellular adhesion molecule‐1 (ICAM‐1, CD54) and L ‐selectin have been reported to increase in patients with allergic diseases and asthma. The present study was therefore undertaken to investigate the expression of CD69, CD54, and L ‐selectin by T lymphocytes of children with asthma, before and after immunotherapy. Eighteen children newly diagnosed with asthma, 11 good and nine poor responders to immunotherapy, and 16 normal subjects, were enrolled in this study. The percentages of CD69⁺, CD54⁺, and CD62L⁺ cells in T lymphocytes were measured by using flow cytometry. The levels of CD69, CD54, and CD62L in serum and culture supernatants were determined by using enzyme‐linked immunosorbent assay (ELISA). The expression of CD69 and CD54 on CD3⁺ T lymphocytes was significantly higher in children with asthma than in control patients. All the patient groups expressed (spontaneously and following stimulation with phorbol myristate acetate and ionomycin together with mite‐extract proteins) greater amounts of CD69 and CD54 than did control subjects. With long‐term immunotherapy, the percentages of CD69⁺ and CD54⁺ T lymphocytes were significantly lower in patients with a good response to immunotherapy. Our results also showed significantly lower serum L ‐selectin levels following immunotherapy. In conclusion, successful immunotherapy resulted in decreased expression and production of CD69 and CD54. These results may explain, in part, the clinical efficacy of immunotherapy.     

外周血及移植肾内嗜酸性粒细胞变化的临床意义

为了解移植肾在急性排斥时外周血和移植肾内嗜酸性粒细胞（ＥＯ）变化的意义，动态观察３１例同种异体肾脏移植病人的外周血和移植肾内ＥＯ的变化。结果发现在急性排斥反应时，移植肾内ＥＯ数＞２％者占８０．９％，明显高于肾功能稳定时，Ｐ＜０．０１；重度排斥中血ＥＯ数＞４％者占８２．６％，明显高于肾功能稳定时和中度以下排斥者，Ｐ＜０．０１。结果认为，测定移植肾内的ＥＯ变化可以做为监测急性排斥反应的可靠指标，外周血中的ＥＯ明显增多常提示排斥反应较为严重。     

Species differences in platelet aggregation induced by platelet-activating factor (PAF).

Species differences in platelet aggregation induced by platelet-activating factor (PAF) were investigated by using the same procedure of platelet preparation and biological assay. Washed platelets of six different species (horses, dogs, rats, rabbits, sheep and guinea pigs) were prepared employing the same method and platelet aggregation was induced by C16-PAF. Horse platelets were most sensitive to PAF (8.0 x 10(-12) M) and rabbit platelets activated by 5.0 x 10(-11) M PAF were also sensitive enough to detect PAF in clinical samples.     

红细胞同工酶ADA－EAP一AK_1同步电泳分型方法的建立

综合、改良了分别分型的方法，建立了红细胞同工酶ＡＤＡ一ＥＡＰ一ＡＫ１同步电泳分型方法，为ＡＤＡ，ＥＡＰ和ＡＫ１３种红细胞同工酶在法医学鉴定中更广泛的应用，提供了一种可靠、经济、实用的方法。     

Cloning of Human α-defensin-1 (HNP-1) Gene and Construction of Its Eukaryotic Expression Vector

1 Introduction Defensins are small cationic antimicrobial peptides that function in the host's innate immune system. The human defensin family includes three small peptides from the azurophil granules of polymorphonuclear cells named human neutrophil peptide (HNP)-1, HNP-2, HNP-3,which consist 5%-7% of the protein of human neutrophil. HNP-4 is approximately one hundred times less abundant. They demonstrate antibacterial, antifungal and antiviral properties in vitro. HNPs are important component of nonoxidative mechanism in macrophages, and can direct inactivate the enveloped viruses. Because only special cells express defensins. And it is hard to extract them naturally and the production is few. So researcher expect to obtain defensins highly through heterogenous expression by gene engineering technology. In order to express HNP-1, we cloned the cDNA of HNP-1 from human polymorphonuclear cells in peripheral blood, and constructed its eukaryotic expression vector, which provided a base for the further study on its mechanism of antimicrobial effect.     

抗人喉癌/抗VEGF双功能抗体制备及应用研究

目的：研制抗人喉癌／抗血管内皮因子（VEGF）双功能克隆抗体，用于喉癌抗血管生成治疗。方法：采用二次杂交瘤技术制备抗人喉癌／抗VEGF双功能抗体。经酶联免疫吸附试验法和SP法检测喉癌及癌前病患者血清及癌组织中VEGF的含量表达。结果：获得6株分泌抗人喉癌／抗VEGF双功能抗体的杂交瘤，经免疫组化证实与喉癌细胞特异性结合率为93％，而与血管内皮细胞结合率为89％。血清中VEGF含量表达，喉癌组与癌前病组及正常对照组相比差异均显著。IgG亚型鉴定为IgG2aBSAb抗体效价为1：25 600倍（ELISA法）。结论：二次杂交瘤法制备的双功能抗体具有均匀性、可控性、效价高、稳定性好，可用于喉癌抗血管生成治疗，动态检测可作为判断喉癌预后的客观指标。     

Role of a conserved amino-terminal sequence in the ecotropic MLV receptor mCAT1

The amino-terminus of mCAT1 and homologous proteins is predicted to form a positively charged, amphipathic alpha helix on the cytoplasmic side of the plasma membrane. Peptides with similar sequence motifs often provide membrane anchors, protein-protein interaction domains, or intracellular transport-targeting signals. Deleting most of the cytoplasmic N-terminal sequence of mCAT1 led to reduced expression on the cell surface and accumulation in the endoplasmic reticulum but did not abrogate receptor function. Surprisingly, when the N-terminal 36 or 18 amino acids of mCAT1 were fused to green fluorescent protein (gfp), gfp accumulated almost exclusively in mitochondria. Mitochondrial targeting depended on arginines at positions 15 and 16 and was inhibitable by downstream transmembrane sequences. Although the full-length mCAT1 was not detected in mitochondria, the mitochondrial-targeting property of the N-terminal sequence fused to gfp is conserved in orthologous and paralogous proteins that diverged approximately 80 million years ago, suggesting a conserved biological function. We propose that the conserved N-terminal motif of CAT proteins provides a regulatable signal for transport to, or retention in, different cell membrane compartments.     

Apolipoprotein D is down-regulated during malignant transformation of neurofibromas

Apolipoprotein D (apoD) expression was studied in nonneoplastic peripheral nerve, neurofibromas (NFs), and malignant peripheral nerve sheath tumors (MPNSTs) by quantitative polymerase chain reaction, in situ hybridization, and immunohistochemistry. Multiplex quantitative polymerase chain reaction for messenger RNA was performed on a series of formalin-fixed and paraffin-embedded specimens that included 9 MPNSTs, 12 NFs, and 4 normal peripheral nerves. The average apoD expression was 108-fold decreased (DeltaCt = -7.3) in the MPNSTs compared with the NFs (P < .05). ApoD expression levels were 3.0-fold elevated (DeltaCt = 1.7) in the NFs compared with nonneoplastic peripheral nerve (P < .05). In situ hybridization for apoD RNA was performed on a separate series of 10 cases in which each microscopic section included both MPNST and the NF from which it arose. These studies confirmed elevated apoD expression in NFs compared with MPNSTs and demonstrated that this expression was variable among individual cells within the NFs. Differential expression by immunohistochemistry could only be demonstrated in selected areas, most likely because apoD protein is a small molecule that is secreted out of the cell into the extracellular space and plasma. ApoD expression initially increases a small amount with the formation of NFs from nonneoplastic peripheral nerve and subsequently decreases markedly as NFs transform into MPNSTs. This expression pattern may serve as a marker for cell cycle inhibition during peripheral nerve tumorigenesis.