免疫磁珠分离技术纯化和检测泰泽氏病原体的研究

目的建立泰泽氏病原体(Ty)纯化方法,获得纯的菌体供抗原研究,为ELISA诊断抗原的制备提供依据;并尝试建立Ty隐性感染血清学抗原检查方法。方法选择特异性抗体包被磁珠,从感染大鼠肝脏中富集和纯化Ty;用SDS-PAGE和Western blot技术考察纯化Ty的蛋白和抗原图谱;同时用免疫磁珠分离技术(IMS)直接检查隐性感染大鼠肠道上皮细胞内的Ty。结果用辛酸-硫酸铵纯化的Ty单克隆抗体M5以0·5μg/107beads以上浓度包被抗IgG抗体预结合的磁珠4h,可最大效率地富集Ty;分离反应进行1h,敏感性达到103菌体/mL;吖啶黄染色镜检法可以直接、快速地观察到结合于磁珠上的细菌;抗原分析表明,IMS较好地去除了肝组织和真核细胞成分,纯化的Ty RJ株具有3个免疫优势的抗原成分,相对分子质量(Mr)分别为160×103、116×103、55×103;此外,IMS法可直接从隐性感染大鼠盲肠上皮细胞中检测到少量寄生的Ty。结论用IMS技术可有效地富集和纯化Ty,并可以作为Ty隐性感染血清学抗原检查的候选方法。     

药用胶带纸层厚度检测系统

设计并试制了一种药用胶带纸层厚度在线检测系统．该系统将高精度电容传感器与计算机结合起来，同时配有丰富的外围器件，实现了多种实际生产中所需要的功能．它的出现，彻底解决了药用胶带纸胶层厚度在线检测这个难题，经过两年多的实际应用的检验，该系统实现了在线非接触测量，精度高，动态性能好，实现功能全面，价格合理，是药用胶带纸生产的必备检测仪器．由于该系统还可以很方便在纸张、塑料薄膜等其它生产线上进行推广，因而，其应用前景十分看好．     

Prevalence of latex sensitization and allergy in Portuguese children

The prevalence of latex allergy has been increasing not only in risk groups but also in the general population, where it is accepted to average 1%. In children, latex sensitization prevalence studies are scarce and involve different population sampling and allergy testing methods, which makes it difficult to compare across studies. Nevertheless, existing studies point towards a low prevalence of latex allergy in children, which still needs to be confirmed in the Portuguese population. Aiming at studying the prevalence of latex sensitization and allergy in a sample of Portuguese children, we studied 182 children from two different hospital outpatient clinics. A standardized questionnaire focusing on atopic background, previous history and allergic signs or symptoms on exposure to latex or fruits was given to all children and parents. Skin prick testing was performed with a battery of common aeroallergens as well as latex. Serum total IgE, Phadiatop, F x 5E and latex-specific IgE were determined in all children. Specific IgE to latex-crossreacting fruits was determined in latex-sensitized children. Based upon the questionnaire, the prevalence of latex allergy would be 0.5%. The prevalence of latex sensitization would be 3.8%, when based solely upon skin prick testing, and 12.1% (>/=0.35 IU/ml) or 6.6% (>/=0.70 IU/ml) when based singly upon determination of latex-specific IgE. When positive results for either test were considered, the prevalence of latex sensitization was 14.3%. All latex-sensitized children were atopic. Sensitivity to latex-crossreacting foodstuffs was demonstrated in 61.5% of latex-sensitized children (16/26). This study shows that the prevalence of latex allergy and sensitization in Portuguese atopic and non-atopic children, as analysed using various diagnostic methods, is similar to that observed in other countries. In addition, the assessment of latex allergy and sensitization should always include skin prick testing and determination of serum IgE.     

The in vitro elution characteristics of vancomycin and tobramycin from calcium sulfate beads

The purpose of this study was to determine the elution characteristics of vancomycin and tobramycin when mixed with calcium sulfate to form antibiotic beads. Calcium sulfate was combined with vancomycin and tobramycin separately to form 2 types of antibiotic beads, which were packaged and labeled separately. The packaged calcium sulfate beads with vancomycin and tobramycin were then gas sterilized. The beads were placed in phosphate-buffered saline and kept at 36 degrees C for 6 weeks. Two separate series of assays were run simultaneously for both types of beads. In one assay, a bead containing vancomycin was placed in a fresh vial of phosphate buffered saline after each assay. The same was done with beads containing tobramycin. In the second series of assays, 9 vials of phosphate buffered saline each containing 1 vancomycin bead and 9 vials of phosphate buffered saline each containing 1 tobramycin bead was arranged. The phosphate-buffered saline was then assayed at predetermined times for both the vancomycin bead series and the tobramycin bead series. The amount of vancomycin and tobramycin assayed nearly equaled the calculated amount of antibiotic per bead measured before bead construction. Also, the elution of antibiotic from the calcium sulfate was complete within 72 hours. In conclusion, the construction and gas sterilization of calcium sulfate beads containing vancomycin and tobramycin does not destroy vancomycin and tobramycin. Also, the complete elution of available vancomycin and tobramycin in calcium sulfate beads occurs within 72 hours.     

重氮乳胶反向凝集试验与乳凝试验在血吸虫病临床诊断中的应用研究

本文应用日本血吸虫虫卵单克隆抗体及虫卵可溶性抗原，分别致敏新型免疫微球载体重氮基聚苯乙烯乳胶，制备出血吸虫反向乳凝试剂与乳凝试剂，并用于检测血吸虫病患者血清标本中的相应抗原和相应抗体，以此试验检测１９７份急慢性血吸虫病患者血清标本。结果反向乳凝试剂检出率为９０．４％，乳凝试剂检出率为９５．２％，均高于反向间接血凝法的８５．０％。反向乳凝试验与乳凝试验联合应用，既可早期诊断，又可判断疗效。试验采用玻片凝集法，３ｍｉｎ内即出结果，方法敏感．特异、简便和快速。     

Latex allergy in infants younger than 1 year

BACKGROUND: The prevalence of latex allergy in children is increasing worldwide. Previous multiple operations or atopic predisposition are known risk factors. In contrast, only sporadic cases of latex allergy have been reported in infants younger than 1 year, and the causative latex-containing products or symptoms in young infants have not been studied in detail. OBJECTIVE: The purpose of this study is to analyse the symptoms and risk factors of latex allergy in young infants. METHODS: Cases of latex allergy in infants younger than 1 year were studied in detail. Clinical course, causative latex-containing products were spotted and detailed analysis for latex allergy in patients and patients' parents was performed. CONCLUSION: We report nine cases of latex allergy in infants younger than 1 year. None of them have any abnormality or previous operations. Six patients had atopic eczema/dermatitis syndrome, one patient had bronchial asthma, whereas two patients had no overt allergic diseases. Symptoms of latex allergy were wheezing, swelling of face or lips, facial rash, or anaphylaxis, and causative latex-containing products were teat, pacifier, nose cleaner, teether, balloon, or enema tube. All of the nine patients had positive skin prick test to latex and extract from causative latex-containing products, whereas eight patients had positive serum latex-specific IgE. Study for family history revealed that latex allergy was noted in either father or mother in six patients, in both father and mother in one patient, whereas no latex allergy was noted in parents in two patients. It should be noted that all of these patients had latex-induced symptoms at home. Latex allergy in young infants may not be unusual. Physicians should be aware of latex allergy, and care should be taken to avoid contact with latex in young infants, especially when there is family history for latex allergy.     

一种LDL吸附剂载体-聚丙烯酰胺微球的合成及应用

研究用于低密度脂蛋白 (L DL)吸附的聚丙烯酰胺微球载体的合成工艺、结构特性及吸附 L DL 的性能 ,为进一步研发 L DL 吸附剂载体提供实验依据。采用反相悬浮聚合法按一定的配方合成聚丙烯酰胺微球载体 ,通过扫描电镜、图像分析仪、X光小角散射等手段对其结构特性 (粒径、孔径等 )进行表征 ;同时在微球上固定丝氨酰 -天冬氨酰 -谷氨酸 (SDE)三肽配体制成 L DL 吸附剂 ,通过体外静态吸附对其吸附性能进行了初步研究。结果表明微球粒径为 14 2 .1μm,孔径为 119.8nm,符合作为 L DL 吸附载体的需要 ;在交联剂与单体总量一定的条件下 ,微球孔径随着交联剂用量的增加而减小 ;合成的聚丙烯酰胺微球对 L DL 的非特异性吸附很小 ,而在其上偶联配体制成吸附剂后 ,又表现出对 L DL 的特异性吸附。本实验合成的聚丙烯酰胺微球是一种有效的 L DL 吸附剂载体。     

The hevein domain of the major latex-glove allergen Hev b 6.01 contains dominant T cell reactive sites

BACKGROUND: Sensitization to natural rubber latex (Hevea brasiliensis) is a major cause of occupational asthma and rhinitis affecting frequent latex-glove users. Hev b 6.01, a known major latex allergen, is cleaved naturally into hevein (4.7 kDa) and a C-terminal fragment (14 kDa). Hevein is an abundant protein in latex-glove extracts. As the immune response to allergens is initiated by activation of allergen-specific CD4(+) T cells, identification of dominant T cell epitopes is crucial for the development of specific immunotherapy. OBJECTIVE: To identify dominant T cell epitopes of Hev b 6.01 in latex-allergic glove users. METHODS: Ten latex-allergic frequent glove users and six non-latex-allergic atopic control subjects were selected, based on clinical symptoms and positive latex-specific serum IgE. Serum IgE reactivity to glove extract and recombinant Hev b 6.01 (rHev b 6.01) were analysed by ELISA. Latex-specific short-term oligoclonal T cell lines were generated from peripheral blood of latex-allergic subjects. These lines were tested for proliferative responses to overlapping 20-mer peptides of the Hev b 6.01 molecule. CD4(+) T cell intracellular cytokines, IL-4 and IFN-gamma were assessed following stimulation with immobilized anti-CD3 in the presence of IL-2. RESULTS: All ten of the latex-allergic patients showed serum IgE binding to glove extract while eight of these also showed IgE binding to rHev b 6.01 by ELISA. Western blotting confirmed reactivity with rHev b 6.01 at around 20 kDa. T cell proliferation assays showed that latex-specific T cell lines from all subjects responded to one or more peptides, with greatest frequency of reactivity to peptides Hev b 6.01 p(10-29) and Hev b 6.01 p(19-38) in the hevein domain. An allergic-type cytokine profile with considerable IL-4 in addition to IFN-gamma was evident from intracellular cytokine staining. CONCLUSION: Hevein is an important T cell as well as B cell immunogen and contains dominant T cell reactive sites.