肿瘤坏死因子 α抑制剂对妊娠的影响

 肿瘤坏死因子 α（TNF-α）是类风湿关节炎（RA）发病和维持关节慢性滑膜炎症反应的最重要的致炎性细胞因子之一，它在 T 细胞依赖的炎症性肠病（IBD）发病机制中也起着十分重要的致炎作用。大量临床研究证实，TNF-α抑制剂（TNFAI）可改善 RA 患者关节功能，减少 RA临床活动性，并延缓关节损坏的进展^[1]。各种 TNFAI 也逐渐用于治疗其他风湿性疾病，如银屑病、幼年特发性关节炎、强直性脊柱炎等。某些 TNFAI 被证明对难治性 IBD 有效。目前经美国 FDA 批准上市的 TNFAI 类药物共有 3 种，它们是依那西普（Etanercept，商品名Enbrel），英利西单抗（Infliximab，商品名 Remicade）和阿达木单抗（Adalimumab，商品名Humira）。依那西普是 II 型TNF受体（TNFR-II）与 IgG1-Fc 的融合蛋白，用药方式为皮下注射。英利西单抗是由人 IgG1-Fc 和鼠 Ig 可变区组成的嵌合体 TNF-α 单抗，阿达木单抗是人源 IgG1 型 TNF-α 单抗，这2 种药物均经静脉注射给药[1]。TNFAI的主要不良反应有注射部位反应、感染、肿瘤、淋巴增殖性疾病、神经脱髓鞘病变以及狼疮样综合征^[1]。由于风湿性疾病多累及生育年龄女性，同时以 TNFAI 为代表的生物制剂应用越来越广泛，TNFAI 对妊娠是否有影响日益受到临床关注。我们复习了2007 年 2 月1日以前 PubMed 中关于上述 3 种 TNFAI对妊娠影响的临床观察文献，现综述如下。......     

Inhibitory effect of antiserum to surface antigen P50 of Babesia gibsoni on growth of parasites in severe combined immunodeficiency mice given canine red blood cells

The inhibitory effect of an antiserum to surface protein P50 of Babesia gibsoni on the growth of the parasite was determined with severe combined immunodeficiency mice given canine red blood cells. The antiserum to the recombinant P50 protein significantly inhibited the parasite growth, indicating that P50 might be a useful vaccine candidate.     

胰腺导管腺癌和慢性胰腺炎中18q21杂合性缺失及相关性分析

目的 探讨18q21在人胰腺导管腺癌和慢性胰腺炎中杂合性缺失(LOH)的情况及其相关因素.方法 选择18q21上的位点RP11-729G3和RP11-850A17作为目的 片段,选择接近18号染色体着丝粒的位点RP11-621L6作为参照位点,利用细菌人工染色体(BAC)提取、纯化相应位点的DNA,用切口平移法分别标记生物素和地高辛后制成双色探针,应用荧光原位杂交技术(FISH)检测30例胰腺导管腺癌和10例慢性胰腺炎石蜡包埋组织切片中18q21 LOH情况,并收集、整理相应临床病理资料进行相关性分析.结果 30例胰腺导管腺癌中在RP11-729G3位点有25例(83.3%)有LOH,在RP11-850A17位点26例(86.6%)有LOH,其中25例在RP11-729G3和fuP11-850A17两个位点均有LOH,1例仅在RP11-850A17位点有LOH.10例慢性胰腺炎中均未发现18q21 LOH.经统计学分析发现,18q21 LOH在慢性胰腺炎和胰腺导管腺癌中差异有统计学意义(P<0.01),且RP11-729G3和RP11-850A17两个位点LOH有高度相关性(Phj系数=0.877),但和临床病理各因素间未发现明显相关.结论 18q21 LOH在胰腺导管腺癌中属于高频事件,并且位点RP11-729G3和RP11-850A17的LOH有高度相关性,可能在胰腺导管腺癌发生发展中起重要作用.在临床诊断中18q21LOH也能作为较特异的标记辅助诊断.     

Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

In this study, we characterized a Babesia equi Be158 gene obtained by immunoscreening a B. equi cDNA expression phage library with B. equi-infected horse serum. The Be158 gene consists of an open reading frame of 3,510 nucleotides. The recombinant Be158 gene product was produced in Escherichia coli and used for the immunization of mice. In Western blot analysis, mouse immune serum against the Be158 gene product recognized 75- and 158-kDa proteins from the lysate of B. equi-infected erythrocytes. In an indirect fluorescent-antibody test with the mouse immune serum, the Be158 antigen appeared in the cytoplasm of Maltese cross-forming parasites (which consist of four merozoites) and was located mainly in the extraerythrocytic merozoite body. When the recombinant Be158 gene product was used in an enzyme-linked immunosorbent assay as a serological antigen, it was found to react to B. equi-infected horse sera, indicating that the Be158 gene product is useful as a serologically diagnostic antigen for B. equi infection.     

Expression of Babesia equi merozoite antigen 1 in insect cells by recombinant baculovirus and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay

The gene encoding the entire Babesia equi merozoite antigen 1 (EMA-1) was inserted into a baculovirus transfer vector, and a recombinant virus expressing EMA-1 was isolated. The expressed EMA-1 was transported to the surface of infected insect cells, as judged by an indirect fluorescent-antibody test (IFAT). The expressed EMA-1 was also secreted into the supernatant of a cell culture infected with recombinant baculovirus. Both intracellular and extracellular EMA-1 reacted with a specific antibody in Western blots. The expressed EMA-1 had an apparent molecular mass of 34 kDa that was identical to that of native EMA-1. The secreted EMA-1 was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA differentiated B. equi-infected horse sera from Babesia caballi-infected horse sera or normal horse sera. The ELISA was more sensitive than the complement fixation test and IFAT. These results demonstrated that the recombinant EMA-1 expressed in insect cells might be a useful diagnostic reagent for detection of antibodies to B. equi.     

Wnt2信号通路重要成员在胶质瘤中的表达及其意义

近来发现Wnt信号通路对个体发育和肿瘤生成具有重要作用[1-2].为进一步探讨Wnt通路在胶质瘤中是否存在表达异常及其在胶质瘤生成中的作用,我们采用RT-PcR、组织芯片免疫组织化学染色以及蛋白印迹法,对人脑胶质瘤Wnt2通路相关因子的表达变化及其意义进行了分析.     

Immunization with recombinant surface antigen P50 of Babesia gibsoni expressed in insect cells induced parasite growth inhibition in dogs

This is a report of a vaccine trial directed against Babesia gibsoni infection in dogs with the use of the recombinant antigen P50. Dogs immunized with P50 showed partial protection manifested as a significantly low level of parasitemia. The results indicated that P50 is a primary vaccine candidate molecule against canine B. gibsoni infection.     

Effects of hypoxia and tumor necrosis factor-alpha on production of plasminogen activator inhibitor-1 in human proximal renal tubular cells

Chronic hypoxia has been newly proposed as a common mechanism of tubulointerstitial fibrosis in the progression of various chronic inflammatory renal diseases, where PAI-1 plays an important role in the accumulation of extracellular matrix (ECM) through inhibition of plasmin-dependent ECM degradation. In the present study, we investigated the presence of PAI-1 in renal tubular cells by immunostaining renal biopsy samples. We also closely examined the effects of hypoxia and TNF-alpha on PAI-1 expression in cultured human proximal renal tubular cells (HRCs). Confluent cells growth-arrested in DMEM for 24h were exposed to hypoxia (1% O2) and/or TNF-alpha at 10 ng/ml for 24 hours. Amounts of PAI-1 protein and mRNA after stimulation were measured by ELISA and TaqMan quantitative PCR, respectively and compared to those in cells incubated under control conditions (18% O2 without TNF-alpha). HIF-1alpha was demonstrated by immunoblot analysis. In crescentic glomerulonephritis, clusters of proximal tubules were specifically stained for PAI-1. Treatment of 24 hours with hypoxia, TNF-alpha and their combination induced a 2.7-fold, a 1.8-fold, and a 4.6-fold increase in PAI-1 protein secretion, and produced a 3.6-fold, a 3.3-fold, and a 12.1-fold increase at the PAI-1 mRNA level, respectively. Immunoblot analysis revealed that hypoxia-inducible factor-1alpha (HIF-1alpha) was markedly accumulated in the nuclear fraction after 16-hours exposure of HPTECs to hypoxia but not to TNF-alpha. In conclusion, hypoxia induces PAI-1 expression via remarkable nuclear accumulation of HIF-1alpha in HRCs. TNF-alpha can enhance this hypoxia-induced PAI-1 expression.     

Improved enzyme-linked immunosorbent assay using C-terminal truncated recombinant antigens of Babesia bovis rhoptry-associated protein-1 for detection of specific antibodies

An enzyme-linked immunosorbent assay (ELISA) based on a recombinant rhoptry-associated protein-1 (RAP-1) of Babesia bovis has been previously developed, but it was imperfect because some cross-reactions were still present in Babesia bigemina-infected bovine sera. To improve its accuracy for the specific detection of the antibodies to B. bovis, we constructed three C-terminal truncated recombinant antigens of the RAP-1-rCT1 (amino acids [aa] 301 to 408), rCT2 (aa 388 to 490), and rCT3 (aa 466 to 565)-by using a baculovirus expression system and evaluated their diagnostic potentials using ELISA. rCT1 and rCT2 were better diagnostic antigens in their sensitivities and diagnostic efficiencies than rCT3, although none of the recombinant antigens showed any cross-reactivity to B. bigemina-infected bovine sera. These results confirmed that the N-terminal 300-aa region caused cross-reactivity of the entire RAP-1 antigen, and the C-terminal truncated recombinant antigens were shown to be useful reagents for species-specific serodiagnosis.     

Heterogeneity of feline herpesvirus type 1 strains

Summary Heterogeneity of 9 feline herpesvirus type 1 (FHV-1) strains consisting of the prototype C27 strain, one French isolate, six Japanese isolates, and the attenuated vaccine F2 strain was examined by biological, immunological, and molecular biological methods. No significant difference was observed in virus growth and antigenic properties among the strains in Crandell feline kidney cell cultures. Hemagglutination activity was also detected in all extracts of cells infected with each strain. However, in immunoblot analysis, a virus-structural immunogenic protein with an M_r of 36 kDa was lacking in 2 strains, one of which was the vaccine F2 strain, whereas the other immunogenic proteins including three kinds of major glycoproteins were detected in all strains without differences in electrophoretic mobilities. Furthermore, when restriction endonuclease analysis was performed to examine the genomic heterogeneity of strains, the cleavage patterns with the enzymeMluI showed a genomic heterogeneity between wild and vaccine strains. In contrast, only a slight variation in the sizes of some fragments was shown with most of the 7 other enzymes used. These results indicated that the lack of the 36 kDa protein and theMluI cleavage pattern could be used as markers of the vaccine F2 strain. The specific markers are important not only to control the quality of the vaccine but also to evaluate the vaccine immunity in FHV-1 infection in cats.