抗人C1—抑制物单链抗体的构建

由一株抗人Ｃ１－抑物单克隆抗体鼠杂交瘤细胞提取总ＲＮＡ，合成２对与免疫蛋白可变区ＦＲ１和ＦＲ４互补的通用引物，经ＲＴ－ＰＣＲ扩增出抗体的重链可变区和轻链可变区基因。     

肺腺癌细胞微环境及转移与黏附分子表达的关系

目的研究肺腺癌细胞生长环境及转移性与黏附分子CD44v6和CD29的表达关系。方法将起源相同、转移性不同的两个肺腺癌细胞系AGZY和Anip分别用简便肿瘤多细胞球体(MTS)培养法培养，并设常规单层贴壁细胞培养对照。通过倒置显微镜、扫描及透射电镜观察MTS形成情况，并用免疫组化法分别对MTS及贴壁细胞上CD44v6和CD29表达进行检测。结果MTS培养成功，贴壁细胞与MTS在细胞结构及细胞连接结构上相似，两种MTS在形态及结构上差异无显著性。免疫组化结果显示，CD29在高转移性的Anip细胞及其MTS上呈阳性表达；在低转移性的AGZY细胞及其MTS上阴性表达。CD44v6在Anip和AGZY细胞及MTS上均呈阳性表达，差异无显著性。贴壁细胞与MTS上两种黏附分子表达均无差异。结论成功建立了一种简易制备MTS的方法。细胞生长方式(单层贴壁与MTS)可能不影响CD44v6和CD29的表达。CD29表达可能与肺腺痛转移性相关；CD44v6表达可能与肺腺癌转移无关。     

腰骶部SPR术中脊神经前后根定位的应用解剖

目的：为ＳＰＲ提供可靠的术中脊神经前、后根鉴别的解剖学依据。方法：在２０例成人脊柱标本上,去除后部结构,暴露整个马尾神经,对Ｌ１～Ｓ２节段的前后根进行形态学观察和测量。结果：脊神经后根位于马尾的后半部,前根则位于前半部。脊神经后根较相应的前根粗大,后根从Ｌ１～Ｓ１逐渐增大,以Ｓ１为最粗大;前根则以Ｌ３最粗大。相应前后根出硬脊膜前,有一段相互贴附并紧贴硬脊膜侧壁。结论：在多椎板切除ＳＰＲ术中前、后根的定位及鉴别,暴露时可根据前、后根出硬脊膜前的相互贴附;在限制性椎板切除时则可通过脊髓外侧索和Ｌ１前、后根之间的最下端的齿状韧带加以鉴别     

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。     

降钙素基因的化学合成与克隆

本文报道用化学法全合成降钙素的基因。基因全长113碱基对。采用大肠杆菌偏爱密码子。整个基因分为6个片段合成。用T~4DNA连接酶一次连接成功,序列分析证明合成的基因序列与设计的一致。     

用BA－ELISA斑点法检测痢疾杆菌免疫小鼠GALT中的特异性抗体分泌细胞（ASC）

改进的ＢＡ－ＥＬＩＳＰＯＴ法使免疫酶斑更为清晰，保存时间延长。用此法检测了痢疾杆菌福氏２ａ经口及腹腔免疫后，小鼠派伊尔氏（ＰＰ）淋巴结、肠系膜淋巴结（ＭＬＮ）及脾脏（ＳＰＬ）中特异性ＩｇＡ、ＩｇＧ、ＩｇＭ抗体分泌细胞（ＡｎｔｉｂｏｄｙＳｅｃｒｅｔｉｎｇｃｅｌｌ，ＡＳＣ）的动态变化，得到有规律的结果：两种免疫途径均能在ＰＰ及ＭＬＮ中诱导出３类特异ＡＳＣ的显著升高，但口服导致的升高其持续时间较腹腔途径为短。此外，腹腔途径还能诱导ＳＰＬ中３种ＡＳＣ升高。     

Commercial preparations of lipoteichoic acid contain endotoxin that contributes to activation of mouse macrophages in vitro

Lipoteichoic acids (LTA), cell wall components of gram-positive bacteria, have been reported to induce various inflammatory mediators and to play a key role in gram-positive-microbe-mediated septic shock. In a large number of these studies, investigators used commercially available LTA purified from a variety of gram-positive bacteria, including Staphylococcus aureus, Bacillus subtilis, and Streptococcus sanguis. We report here that, although these commercially available LTA could be readily shown to stimulate production of nitric oxide (NO) in RAW 264.7 mouse macrophages, the activity was dramatically inhibited by polymyxin B, a relatively specific inhibitor of endotoxin biological activity. One-step purification of the commercially available S. aureus LTA using hydrophobic interaction chromatography resulted in two well-separated peak fractions, one highly enriched for LTA and a second highly enriched for endotoxin. The LTA-enriched fractions did not induce production of NO in RAW 264.7 macrophages, although they caused a dose-dependent induction of NO in the presence of low concentrations of gamma interferon (IFN-gamma) (which by itself induced little NO), regardless of the presence of polymyxin B. In contrast, the endotoxin-enriched fractions by themselves inhibited in high levels of NO in RAW 264.7 macrophages but activity was almost completely inhibited in the presence of polymyxin B. Consistent with these findings, our data also indicate that commercial LTA preparations from S. aureus, B. subtilis, and S. sanguis were not able to induce NO from lipopolysaccharide-hyporesponsive C3H/HeJ mouse peritoneal macrophages, but in the presence of IFN-gamma, these LTA preparations were able to induce relatively high levels of NO from C3H/HeJ macrophages. These results indicate that commercially available LTA can contain contaminating and potentially significant levels of endotoxin that can be expected to contribute to the putative macrophage-stimulating effects of LTA as assessed by NO production. The fact that the purified LTA, by itself, was not able to induce significant levels of NO secretion in RAW 264.7 macrophages supports the conclusion that caution in attributing high-level biological activity to this microbial cell wall constituent should be exercised.     

大鼠肝抑素纯化及其生物活性的检测

用ＳｅｐｈａｄｅｃＧ－５凝胶过滤层析法，进一步纯化具肝抑素生物活性的大鼠肝蛋白质粗提品，以分离的大鼠再生肝的肝细胞为靶细胞，体外检测各洗脱峰浓缩物对肝细胞增殖的制率结果证明，Ｅ峰浓缩物的抑制作用最强，其活性比为粗提品的２０倍，ＳＤＳ聚丙烯酰胺电泳图及蛋白质迁移率测定表明，该浓缩物的主要成分为分子量１３．５ｋＤ的多肽。本研究对大鼠肝抑素做了初步纯化，验证了该物质在肝再生中起重要调控作用的生物效应。     

Role of BLNK in oxidative stress signaling in B cells

BLNK (B cell linker protein) represents a central linker protein that bridges the B cell receptor-associated kinases with a multitude of signaling pathways. In this study, we have investigated the role of BLNK in oxidative stress signaling in B cells. H2O2 treatment of B cells induced a rapid tyrosine phosphorylation of BLNK in a H2O2 dose-dependent manner, which was inhibited in Syk-deficient DT40 cells. Calcium mobilization in BLNK-deficient as well as Syk-deficient and phospholipase C (PLC)-gamma2-deficient cells after H2O2 treatment was completely abolished. These were derived from decreased inositol 1,4,5-trisphosphate generation through PLC-gamma2 in BLNK-deficient cells. Moreover, viability of BLNK-deficient as well as PLC-gamma2-deficient cells after exposure to low doses of H2O2 was dramatically enhanced compared with that of the wild-type cells. Furthermore, c-Jun N-terminal kinase activation following high doses of H2O2 stimulation, but not low doses of H2O2 stimulation, was abrogated in BLNK-deficient as well as Syk-deficient cells. These findings have led to the suggestion that BLNK is required for coupling Syk to PLC-gamma2, thereby accelerating cell apoptosis in B cells exposed to low doses of H2O2.     

Detection and identification ofMycobacterium avium in the blood of AIDS patients by the polymerase chain reaction

One hundred fifty-three blood samples from patients positive for the human immunodeficiency virus (HIV) were analyzed by polymerase chain reaction (PCR) to detect the presence ofMycobacterium avium. Samples were collected from patients who also had blood cultures performed by a radiometric method. Blood samples were centrifuged on a Ficoll-Hypaque gradient to purify peripheral blood mononuclear cells. The purified cells were washed and incubated with a resin, boiled to release mycobacterial DNA, and then amplified. Polymerase chain reaction products were detected by a nonisotopic method. A 123 base-pair (bp) insertion sequence, namely IS6110, fromMycobacterium tuberculosis complex was also included in the reaction as an internal control ofTaq polymerase activity to exclude the presence of enzyme inhibitors. This IS6110 fragment can be distinguished from the 383 bp target product on ethidium bromide-stained agarose gel and may also be used in a colorimetric assay. Such results were compared with the results of culture and indicated that the assay is as sensitive as bacteriological methods, though faster.