白细胞粘附作用对ELISA检测HBsAg干扰现象的探讨

目的 探讨在ELISA检测HBsAg试验中，标本混入WBC后，WBC的粘附现象对试验结果的干扰。方法 观察WBC粘附现象对测定结果的干扰情况，以及血浆中WBC含量与干扰程度的关系，同时采用增加洗涤次数的方法对抗WBC粘附的效果观察。结果 标本混入WBC，WBC在反应板内发生粘附。WBC的中性粒细胞含过氧化物酶对试验结果造成干扰；标本中单位体积WBC含量与WBC的粘附量不成正比，但WBC的有效粘附数与OD值直接相关，粘附量越多，OD值越高；且WBC的粘附经洗涤10次也未能完全洗脱。结论 在ELISA检测HBsAg工作中应注意WBC对试验产生的非特异性干扰，采用多种方法防止WBC的混入。     

Demonstration of adherence properties of Porphyromonas gingivalis outer membrane vesicles using a new microassay

Vesicles made by Porphyromonas gingivalis possess several biological activities, including the ability to adhere to oral surfaces and to bacteria. In this study, a new and simple method was developed to measure the adherence capability of outer membrane vesicles from P. gingivalis . Vesicles were conjugated to fluorescent microspheres (0.7 μ) and added to wells of a Teflon-coated microscope slide previously covered with a variety of soluble ligands. After incubation and washes, the number of fluorescent microspheres per microscopic field were counted. Vesicle-coated microspheres attached best to gelatin (<200 per field), whereas other compounds (such as fibronectin, fibrinogen, collagen and laminin) provided moderate attachment, and no attachment was observed to bovine serum albumin. Adherence to any of the tested ligands was not observed when fluorescent micro-spheres were conjugated to bovine serum albumin or lipopolysaccharides from P. gingivalis. The adherence of vesicle-coated microspheres to ligands was not significantly affected when the pH of the reaction mixture was between 4 and 10. None of the tested carbohydrates lowered the attachment capability of vesicle-coated microspheres to substrates. When vesicle-coated microspheres were treated with trypsin and chymotrypsin or heated, this resulted in a significant loss of attachment, suggesting a possible involvement of proteinaceous molecules in the process. The present study confirms that vesicles of P. gingivalis are capable of attachment to various molecules and indicate their potential role in colonization.     

Adherence of Staphylococcus aureus to cultures of human peritoneal mesothelial cells

Strains of Staphylococcus aureus, isolated from the effluentof patients with peritonitis on CAPD (continuous ambulatoryperitoneal dialysis), adhered well to both cultured human mesothelialcells and to fibronectin, but not to laminin or gelatin. Mesothelialcells grown in medium M199 exhibited more surface fibronectincompared to cells grown in MEM-Dval and demonstrated higherlevels of S. aureus adherence. Soluble fibronectin concentrations up to lOµg/ml increasedthe adherence of S. aureusto cultured mesothelial cells. Thedose-response curve was consistent with the binding of fibronectinto a saturable receptor of apparent dissociation constant (K_D)= 1.7xlO^–10 M. This corresponds closely to the K_D (2xlO^–10M) of the staphylococcal fibronectin-binding protein. S. aureus adherence was increased following the preincubationof mesothelial cell monolayers with interleukin-1 and was maximalafter 6 h preincubation. Treating mesothelial cells with interferon-gammafor 48–72 h reduced the adherence of S. aureus.     

带状疱疹患者红细胞免疫功能的测定

采用红细胞酵母菌花结试验检测了３２例带状疱疹患者的红细胞免疫粘附活性。结果显示：疾病组红细胞Ｃ３ｂ受体花环率（７．７６±１．７６％）较对照组（９．４６±ｌ．７３％）明显降低（Ｐ＜０．０１），而红细胞免疫复合物花环率（９．９２±１．９９％）较对照组（４．７１±２．２８％）明显增高（Ｐ＜０．０１）。     

User satisfaction with a real-time automated feedback system for general practitioners: a quantitative and qualitative study.

OBJECTIVE: The GRIF automated feedback system produces real-time comments on the appropriateness of diagnostic tests ordered by general practitioners (GPs) based on recommendations from accepted national and regional practice guidelines. We investigated the experiences of GPs with this system and, more specifically, with the recommendations produced by the system as well as their views on using this system in daily practice. SETTING: We tested the GRIF system in an experiment in a laboratory setting and in a daily practice trial. STUDY PARTICIPANTS: General practitioners. INTERVENTION: In the laboratory experiment, GPs used the GRIF system to assess the appropriateness of 30 request forms. Each of the GPs was confronted with requests they had submitted to the diagnostic unit of the hospital in the past. In the field trial, the GRIF system was applied during patient consultations for 1 year. MAIN OUTCOME MEASURES: We measured GPs' satisfaction with the system using a questionnaire, and also conducted group discussions (in the laboratory experiment) and in-depth interviews (in the field trial) to elicit GPs' opinions of and experiences with the system. In addition, we explored GPs' reasons for not accepting the comments offered by the GRIF system. RESULTS: The results show that the GPs in the laboratory experiment had more positive attitudes towards the system compared with participants in the field trial. All discussion groups and most of the GPs in the field trial regarded receiving the immediate feedback during the test ordering process as an important advantage. The most frequently mentioned reason to reject the recommendation was disagreement with the content and/or the recommendations in the practice guidelines. CONCLUSION: Apart from securing agreement on guideline content, a prerequisite for using GRIF in daily practice on a large scale is that more attention is paid to promotion of the guidelines and their adoption, and stimulation of a positive attitude towards the practice guidelines among the users.     

The effect of milk and casein proteins on the adherence of Streptococcus mutans to saliva-coated hydroxyapatite

Experiments sought to determine the nature of the binding of milk proteins to hydroxyapatite (HA) and to saliva-coated hydroxyapatite (sHA), and to determine the effect of milk and casein on the adherence of Streptococcus mutans GS-5 to sHA. The binding of radiolabelled -casein to HA was reduced when incubated simultaneously with parotid saliva, and enhanced in the presence of milk. The binding of β- and κ-casein to HA was unaffected by the presence of parotid saliva and enhanced by the presence of milk. The in vitro bacterial adherence of Strep. mutans GS-5 to sHA beads was reduced when beads were coated with milk instead of buffer, or when bacteria were added to sHA in the presence of milk instead of buffer. Casein proteins (, β, κ) added to sHA simultaneously with bacteria inhibited the adherence of Strep. mutans GS-5 to sHA. κ-Casein, when bound to sHA, inhibited streptococcal adherence to sHA; - and β-casein, when bound to sHA, had no effect on streptococcal adherence. Fractionation of κ-casein by anion-exchange chromatography revealed the anti-adherence activity of κ-casein was mediated primarily by a 40,000 mol. wt. glycoprotein-containing fraction. These data show that milk, particularly κ-casein fractions, can modulate the adherence of Strep. mutans GS-5 to SHA surfaces in vitro.     

Self-efficacy and the maintenance of exercise participation in older adults

The role played by exercise self-efficacy in the maintenance of exercise participation of previously sedentary middle-aged adults 4 months after the termination of a formal exercise program is reported. Correlational and multiple regression analyses examined the influence of self-efficacy, physiological (aerobic capacity, sex, body composition), and behavioral (past exercise frequency and intensity) parameters in the maintenance of exercise participation. Self-efficacy significantly predicted exercise behavior at follow-up when controlling for biological and behavioral influences. Aerobic capacity, exercise efficacy, and exercise behavior in combination were significantly related to current energy expenditure in aerobic physical activity. The discussion focuses on the need to examine the impact of different correlates of exercise behavior at different stages of the exercise process.This project was funded by Grant AG07907 from the National Institute on Aging.     

I Get by with a Little Help from my Family and Friends: Adolescents' Support for Diabetes Care

Evaluated and compared the support provided by family membersand friends for adolescents' diabetes care. Family and friendsupport also were examined in relation to other measures ofsocial support, to demographic variables (age, gender, durationof diabetes) and to adherence. Using a structured interview,74 adolescents with diabetes described the ways that familymembers and friends provided support for diabetes management(insulin shots, blood glucose monitoring, eating proper meals,exercise), and for helping them to "feel good about their diabetes."Families provided more support than friends for three managementtasks (insulin injections, blood glucose monitoring, meals);this support was largely instrumental. In contrast, friendsprovided more emotional support for diabetes than families.Greater family support was related to younger age, shorter diseaseduration, and better treatment adherence. Implications of thefindings include encouraging parents to remain involved in adolescents'treatment management, and involving peers as supportive companionsfor meals and exercise.     

Comparison of Different Types of Oral Adsorbent Therapy in Patients with Chronic Kidney Disease: A Multicenter,Randomized, Phase IV Clinical Trial

PurposeOral adsorbents delay disease progression and improve uremic symptoms in patients with chronic kidney disease (CKD). DW-7202 is a newly developed oral adsorbent with high adsorptive selectivity for uremic toxins. We evaluated patient preference for and adherence to DW-7202 versus AST-120 therapy and compared treatment efficacy and safety in patients with pre-dialysis CKD.Materials and MethodsA seven-center, randomized, open-label, two-way crossover, active-controlled, phase IV clinical trial was conducted. Patients with stable CKD were randomly assigned to receive DW-7202 (capsule type) or AST-120 (granule type) for 12 weeks. The groups then switched to the other adsorbent and took it for the next 12 weeks. Patient preference was the primary outcome. Secondary outcomes included changes in estimated glomerular filtration rate (eGFR) and serum creatinine, cystatin C, and indoxyl sulfate (IS) levels.ResultsSignificantly more patients preferred DW-7202 than AST-120 (p<0.001). Patient adherence improved after switching from AST-120 to DW-7202; there was no apparent change in adherence after switching from DW-7202 to AST-120. Changes in eGFR and serum creatinine, cystatin C, and IS levels were not significantly different according to adsorbent type. There was also no significant difference in the incidences of adverse events during treatment with DW-7202 and AST-120.ConclusionDW-7202 can be considered as an alternative to AST-120 in patients who cannot tolerate or show poor adherence to granule type adsorbents. Further studies to evaluate factors affecting patient preferences and improved adherence are warranted (Clinical trial registration No. {"type":"clinical-trial","attrs":{"text":"NCT02681952","term_id":"NCT02681952"}}NCT02681952).     

金黄色葡萄球菌 eap 基因的克隆与表达

 目的 构建携带 eap 基因的原核表达载体，诱导表达具有活性的重组 EAP 融合蛋白。 方法 PCR 法扩增金黄色葡萄球菌基因组 DNA，回收、 纯化的扩增产物与 pMD18-T 载体相连接得重组质粒 pMD18-T-EAP，转化 E.coli BL21（DE3）感受态细胞，酶切鉴定；未酶切组作为对照组重组质粒 pMD18-T-EAP 和 pET28a（+）表达载体分别用 Nde I 和 Xho I 限制性内切酶双酶切、连接，转化 E.coli BL21（DE3）感受态细胞，酶切鉴定；空载体作为对照组。用不同浓度（终浓度 1、2、4、8 mmol/L）和不同诱导时间（1、2、3、4、5、6 h）的异丙基-β-D-硫代半乳糖苷（IPTG）对阳性重组菌进行表达优化，分别取 E.coli上清液和沉淀做电泳分析。应用 MagneHis^TM 蛋白纯化系统纯化重组 EAP 融合蛋白，并通过薄层扫描测定蛋白质的浓度。 结果 所获 eap 基因与 GeneBank 的基因序列同源性 > 99%；氨基酸同源性达 100%。重组质粒经 IPTG 诱导，阳性重组菌转化子均有表达；当吸光度（A ）值等于 0.6 ~ 0.8 时，相对分子质量约 70 000 处出现目的蛋白条带。破碎的重组菌 pET28a-EAP上清液中目的蛋白条带较清楚，沉淀中几乎看不到。终浓度 1 mmol/L 为最佳蛋白表达工作浓度。IPTG 诱导 1 h 重组 EAP 融合蛋白有一定量的表达，随着时间的延长，表达量增加不明显，3 h 时的表达量达最高，之后，蛋白表达量变化不明显。表达的重组 EAP 融合蛋白含量占全菌体蛋白的 29.6%。 结论 成功地克隆和表达了金黄色葡萄球菌重组 EAP 融合蛋白，为进一步研究以 EAP 蛋白作为免疫原预防和治疗由金黄色葡萄球菌引起的疾病奠定基础。