噬菌体表面表达人干扰素αlc/86D

目的为今后干扰素αｌｃ／８６Ｄ突变体文库的建立和筛选高活性的新型干扰素分子打下基础。方法利用ｐｈａｇｅｄｉｓｐｌａｙ技术表达人ＩＦＮαｌｃ／８６Ｄ基因。结果酶联免疫吸附试验、点杂交和Ｗｅｓｔｅｒｎｂｌｏｔ均证明重组噬菌体颗粒具有干扰素的免疫反应性，生物学活性测定表明，当重组噬菌体为５×１０１０ＴＵ时，与４ＩＵ的重组ＩＦＮαｌｂ活性相当。结论人ＩＦＮαｌｃ／８６Ｄ在噬菌体表面能正确折叠和功能性表达。     

类风湿关节炎滑膜组织T7噬菌体展示cDNA文库的构建

目的构建类风湿关节炎(RA)滑膜组织T7噬菌体展示cDNA文库,为筛选和鉴定RA特异性基因的研究奠定基础。方法取确诊RA患者的滑膜组织,用Trizol试剂提取总RNA,Oligotex离心柱分离mRNA,电泳检测其质量,逆转录合成双链cDNA,经末端修平、接头连接、酶切后去除过多接头,收集>300bp的cDNA片段,与T7Select10-3载体连接,体外包装并扩增得到类风湿关节炎滑膜组织T7噬菌体展示cDNA文库。最后,通过铺平板滴度测定及PCR技术鉴定文库质量。结果所构建原始文库重组克隆数为2×107pfu,扩增滴度8.9×1010pfu/ml。PCR法检测片段插入率为90%,插入片段在300～2000bp之间。文库噬菌体裂解液PCR扩增得到BiP基因片段。结论成功构建了高质量的RA滑膜T7噬菌体展示cDNA文库,为下一步RA自身抗原的筛选奠定了基础。     

Elicitation of potent SARS-CoV-2 neutralizing antibody responses through immunization with a versatile adenovirus-inspired multimerization platform

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Screening serum biomarkers for early primary hepatocellular carcinoma using a phage display technique

Hepatocellular carcinoma (HCC) occurs mainly in chronically diseased livers following hepatitis B virus (HBV) or hepatitis C virus (HCV) infection. Early detection and diagnosis of HCC would be of great clinical benefit. In this study, we used a random phage display peptide library and sera from early-stage primary HCC patients (n = 30) to screen potential serum biomarkers for early primary HCC. Age- and sex-matched patients with HBV and/or HCV infection were used as controls. In the screening phase, 19 out of 20 randomly selected phage clones exhibited specific reaction with purified sera IgG from early primary HCC patients, among them 14 coming from the same phage clone with inserted peptidesequence RGWCRPLPKGEG (named HC1). In the validation phase, phage ELISA results showed that the positive reaction rate of the HC1 phage clone was 91.4% with the early HCC group (n = 70), significantly higher than that with the HBV infection group (20.0%) (n = 70), the HCV infection group (12.9%) (n = 70), the HBV + HCV infection group (24.3%) (n = 70), the cirrhosis group (17.1%) (n = 70), and the healthy control group (10.0%) (n = 70). In conclusion, the HC1 mimic peptide showed high diagnostic validity for early primary HCC, and thereby could be a candidate serum biomarker for early primary HCC.     

Effect of ambient light level at the monitor surface on digital radiographic evaluation of approximal carious lesions: an in vitro study

Objectives

This study investigated how ambient light affects the diagnostic accuracy of dental carious lesions on monitors used in dental practice. Specifically, the aim was to evaluate whether a monitor hood for blocking excess ambient light increases practitioners’ ability to accurately diagnose carious lesions on digital radiographs under bright ambient light conditions.

Methods

7 observers evaluated approximal carious lesions on standardized digital radiographs of 100 teeth under 3 ambient light conditions: bright light (> 1000 lx) and dim light (<50 lx) with no monitor hood; and bright light with a hooded monitor. Receiver operating characteristic curves were plotted for all observations. The criterion standard was a histological examination of the teeth. A paired t-test compared the effects of the three lighting conditions. The level of significance was set to p <0.05. Weighted kappa statistics estimated intraobserver agreement.

Results

The diagnostic accuracy for dentine lesions was significantly higher in ambient light<50&hairsp ;lx than on monitors with and without a hood in ambient light>1000 lx. For all observers, diagnostic accuracy of dentine lesions under bright light was higher on a hooded monitor than on a monitor without a hood, but this difference was not significant. Intraobserver agreement varied from moderate to good.

Conclusion

Diagnostic accuracy of those carious lesions that reached into the dentine was significantly higher in ambient light<50 lx than in ambient light>1000 lx. A hooded monitor in bright light was not as effective as a monitor without a hood in dim light.     

全人源抗MLAA-34单链抗体的筛选和鉴定

目的:抗体药物治疗是目前癌症治疗中很有前景的一种方法,利用前期实验得到的MLAA-34纯化蛋白作为抗原,在噬菌体抗体库中进行筛选得到抗MLAA-34的单链抗体,进行测序并鉴定其亲和力。方法:包被纯化的MLAA-34抗原蛋白于96孔板上,封闭过夜,加入1×1012 cfu噬菌体抗体库,4 ℃结合4 h,洗脱,洗脱后的噬菌体感染大肠杆菌TG1扩增,经过3轮淘选,淘选得到的克隆再进行Phage-ELISA的鉴定,筛选得到阳性克隆。结果:纯化的MLAA-34作为抗原,经过在噬菌体抗体库中的3轮固相筛选,噬菌体的回收得到富集,洗脱的噬菌体产率显示回收率逐级增加,富集倍数约1 000倍。最后一轮筛选后挑选出188个重组噬菌体克隆,His标签蛋白作为对照蛋白,应用Phage-ELISA筛选出33个阳性克隆。结论:利用得到的MLAA-34蛋白为目标抗原,对全人源噬菌体抗体库进行淘选,应用Phage-ELISA法挑选出和MLAA-34抗原结合力的阳性噬菌体,为抗MLAA-34抗体的制备奠定了基础。     

Secrets and disclosure in donor conception

This article considers the disclosure, sharing and exchange of information on being donor conceived within families, drawing on data from a study undertaken with donor‐conceived adults registered with UK Donor Link (a voluntary DNA‐linking register). This paper considers the narratives of how respondents found out they were donor‐conceived and what events triggered disclosure of this information. This paper then goes on to examine the role secrecy played in their family life and uses the concept of ‘display’ to explore how secrecy affected their relationships with their immediate and extended family. Secrets are notoriously ‘leaky’ and we found complex patterns of knowing and uncertainty about whom in the family knew that the person was donor‐conceived. We argue that what is kept secret and from whom provides insights into the multifaceted web of social relationships that can be created by donor‐conception, and how knowledge can be managed and controlled in attempts to display and maintain family narratives of biogenetic connection.     

视屏作业人员职业紧张与热应激蛋白70表达关系的研究

目的通过检测视屏作业人员外周血淋巴细胞中热应激蛋白70(HSP70)表达水平,结合职业紧张测试结果进行综合分析,探讨HSP70表达水平作为职业紧张监测生物标志物的可能性。方法采用职业紧张量表(OSI)对宁夏某银行工龄>1a的108名视屏作业人员进行问卷调查,并按照职业紧张因素得分的四分位数间距法将其分为低、中、高3个紧张组,同时应用免疫印迹法(western bolt)测定外周血淋巴细胞中HSP70表达量。结果低、中、高紧张组HSP70表达水平分别为6.20±0.78、5.83±0.62、5.62±0.48,3组间比较,差异有统计学意义(P<0.01);相关分析结果也表明,随着紧张程度的增高,外周血淋巴细胞中HSP70表达水平降低(r=-0.376,P<0.01),且有多个职业紧张因素与外周血淋巴细胞中HSP70水平关系密切。结论外周血淋巴细胞中HSP70表达水平可作为评价慢性职业应激的直接、客观的指标。     

Development of orf virus as a bifunctional recombinant vaccine: surface display of Echinococcus granulosus antigen EG95 by fusion to membrane structural proteins

The parapoxvirus, orf virus (ORFV) causes superficial skin lesions in infected sheep. Unattenuated ORFV is used globally to vaccinate against orf. Recombinant poxviruses are proven delivery systems and we investigated strategies to express the immunogenic Echinococcus granulosus peptide EG95 from ORFV with the aim of developing a recombinant bivalent vaccine. EG95 is an oncosphere protein of the cestode E. granulosus, a parasite responsible for causing cystic hydatid disease in a wide range of hosts including humans and grazing animals such as sheep. Recombinant viruses were produced in which EG95 was expressed by itself or fused to ORFV envelope-associated structural proteins 10 kDa and F1L. Infection studies in sheep showed that specific antibodies were produced against ORFV and EG95 and that the antibody levels against EG95 were comparable to that of animals immunized with purified EG95 in Quil A adjuvant, an immunization regime that is known to afford protection. A single exposure to the dual vaccine has potential for protecting lambs against orf and for priming against EG95 so as to respond strongly to a later injection of EG95 protein.