分析RhE抗原分布检测对临床输血的价值

分析恒河猴E抗原(RhE)分布检测对临床输血的价值,为临床实践提供参考。对本院临床住院及献血人员恒河猴D抗原(RhD)血液样本16 800例进行RhE、e表型鉴定分析,并分析其中2 280例RhE阴性接收RhE阳性血液后不规则抗体的变化,分析患者输血前后血液指标的变化情况。结果显示16 800例RhD血液样本Dee、DEe及DEE表型分别为8 420(50.1%)、6 847(40.8%)和1 533(9.1%);RhE阴性患者接收RhE阳性血液后不规则抗体分析发现产生阳性抗体共有185例,占8.1%,且两组患者输血前后胆红素及网织红细胞无明显变化,而3次及以上输血患者输血后血红蛋白(Hb)及红细胞(RBC)明显升高(P<0.05)。因此,RhE抗原分布检测指导临床同型输血具有重要的价值。     

临床待输血患者RhE抗原和不规则抗体的检测及其临床意义

目的：了解临床待输血患者红细胞Rh血型系统E抗原（RhE抗原）分布,结合抗-E抗体在上述患者中的检出频率及特点,分析常规检测RhE抗原的临床意义。方法：选择待输血患者标本10 000例,采用微柱凝胶法检测RhE抗原,并对患者血液标本进行不规则抗体筛查和鉴定。结果：10000例标本中,RhE抗原阳性率51.46%（5146/10000）,RhE抗原阴性率48.54%（4854/10000）。患者标本不规则抗体筛查阳性共78例,阳性率0.78%（78/10000）;全部检出抗体中,抗-E抗体最多,占23.08%（18/78）,检出的18例抗-E抗体阳性患者中17例有输血史或妊娠史;抗-E抗体在有临床免疫史患者中检出率为0.33%（17/5120）,与无免疫史患者检出率（0.02%,1/4880）比较差异有统计学意义（χ²=13.46,P<0.01）。结论：抗-E抗体是临床输血检测中最常出现的不规则抗体,对待输血患者进行RhE抗原常规检测,实现红细胞RhE血型同型匹配输血,可避免患者产生抗-E抗体,促进临床输血安全。     

Regulatory accepted but out of domain: In vitro skin irritation tests for agrochemical formulations

Several in vitro methods have gained regulatory acceptance for the prediction of skin irritation and corrosion. However, the test guidelines for the majority of in vitro methods do not state whether they are applicable to agrochemical formulations. Hence, we would like to share the results from our routine skin corrosion and irritation testing of agrochemical formulations in which both in vitro (according to OECD TG 431 and OECD TG 439) and in vivo (according to OECD TG 404) tests were conducted as regulatory requirements. The in vitro skin irritation test did not correlate well with the CLP classification by in vivo results (44% sensitivity, 60% specificity, and 54% accuracy, based on 65 data pairs). This indicates a lack of applicability of the current protocol of the in vitro skin irritation test for agrochemical formulations. The data set did not contain formulations which were skin corrosive in vivo and hence its applicability could not be assessed. The correlation of in vitro skin corrosion testing to formulations which were not corrosive in vivo was, however, high (95% specificity based on 81 data pairs).     

Applicability of in vitro tests for skin irritation and corrosion to regulatory classification schemes: Substantiating test strategies with data from routine studies

Skin corrosion or irritation refers to the production of irreversible or reversible damage to the skin following the application of a test substance, respectively. Traditionally, hazard assessments are conducted using the in vivo Draize skin test, but recently in vitro tests using reconstructed human epidermis (RhE) models have gained regulatory acceptance. In this study, skin corrosion (SCT) and irritation tests (SIT) using a RhE model were implemented to reduce the number of in vivo tests required by regulatory bodies. One hundred and thirty-four materials were tested from a wide range of substance classes included 46 agrochemical formulations. Results were assessed according to UN GHS, EU-CLP, ANVISA and US EPA classification schemes. There was high correlation between the two in vitro tests. Assessment of the SCT sensitivity was not possible due to the limited number of corrosives in the data set; SCT specificity and accuracy were 89% for all classification systems. Accuracy (63–76%) and sensitivity (53–67%) were low in the SIT. Specificity and concordance for agrochemical formulations alone in both the SCT and SIT were comparable to the values for the complete data set (SCT: 91% vs. 89% specificity, 91% vs. 89% accuracy and SIT: 64–88% vs. 70–85% specificity, 56–75% vs. 63–76% accuracy).     

Classification and labeling of industrial products with extreme pH by making use of in vitro methods for the assessment of skin and eye irritation and corrosion in a weight of evidence approach

Classification and labeling of products with extreme pH values (?2 or ?11.5) is addressed in chemicals legislation. Following determination of pH and alkaline/acid reserve, additional in vitro tests are needed, especially to substantiate results less than corrosive. However, only limited experience with the practical application of in vitro methods to determine appropriate classifications for pH extreme products is available so far. Expert judgment and weight of evidence are given major roles under the globally harmonized system of classification and labeling of chemicals (GHS) and should be performed on a sound data basis. We have used a tiered testing strategy to assess 20 industrial products (cleaning and metal pretreatment) regarding their corrosive and irritating properties towards human skin models in vitro in the EpiDerm™ skin corrosion and/or skin irritation test. Nine dilutions of individual compounds were additionally tested. Non-corrosive samples were tested in the Hen’s egg test chorioallantoic membrane (HET-CAM). We demonstrate how data is combined in a weight of evidence expert judgment, and give examples of classification decisions. To our knowledge this is the first comprehensive analysis of industrial products with extreme pH values to determine irritating and corrosive properties by making use of in vitro methods in a weight of evidence approach.     

序列特异性引物PCR法检测Rh血型E/e基因型

目的　建立Rh血型E/e基因分型的方法，以检测人群中E/e基因频率。方法　采用快速盐析法抽提样本DNA，采用PCR—SSP方法检测E/e基因。结果　在本研究的RhD阳性人群中E基因频率为0.223，e基因频率为0.777。RhD阴性人群中E基因频率为0.040，e基因频率为0.960。结论　人群中以e基因为主，Rh阳性与阴性人群中E和e基因频率存在明显的差异。     

In vitro human skin irritation test for evaluation of medical device extracts

The aim of this study was to determine if the EpiDerm? reconstructed human skin model (MatTek Corp.) could be an acceptable alternative to the ISO 10993-required rabbit skin irritation test for assessing medical device biocompatibility. Eleven medical device polymers were tested. Four extracts were prepared per polymer, two each with saline and sesame oil; half were spiked with two R-38 irritants, lactic acid for saline extracts and heptanoic acid for the sesame oil extracts. Tissue viability was assessed by MTT reduction and the proinflammatory response was assessed by IL-1α release. LOAELs of 2% for lactic acid in saline and 0.7% for heptanoic acid in sesame oil were determined. A cell viability reduction of >50% was indicative of skin irritation. Cells exposed to saline extracts spiked with 3.25% lactic acid had significantly reduced mean cell viabilities (12.6–17.2%). Cells exposed to sesame oil extracts spiked with 1.25% heptanoic acid also exhibited reduced mean cell viabilities (25.5%–41.7%). All spiked cells released substantial amounts of IL-1α (253.5–387.4 pg/ml) signifying a proinflammatory response. These results indicate that the EpiDerm? model may be a suitable in vitro replacement for the assessment of the irritation potential of medical device extracts.     

RhE相同表型输血与不同表型输血疗效对比观察

目的:比较RhE相同表型输血与不同表型输血的临床效果。方法:选择患者80例,在严密观察患者生命体征的情况下,输注红细胞悬液2个单位,观察2组输血前后血红蛋白(Hb)、血细胞比容(HCT)变化及发生的输血不良反应。结果:输血后相同表型组Hb和HCT均显著高于输血前和不同表型组(P0.05),不同表型组输血前后Hb和HCT,差异均无统计学意义(P0.05),相同表型组输血后仅出现2例发热患者,而不同表型组输血后则以出现血红蛋白尿和腰背痛为主,且两者均显著多于相同表型组(P0.05)。结论:针对存在输血史、妊娠史的患者,应该建议供血机构进行RhE血型鉴定和配型,以便更好的提高输血的有效性和安全性。     

Development of a new in vitro skin sensitization assay (Epidermal Sensitization Assay; EpiSensA) using reconstructed human epidermis

Recent changes in regulatory requirements and social views on animal testing have accelerated the development of reliable alternative tests for predicting skin sensitizing potential of chemicals. In this study, we aimed to develop a new in vitro skin sensitization assay using reconstructed human epidermis, RhE model, which is expected to have broader applicability domain rather than existing in vitro assays. Microarray analysis revealed that the expression of five genes (ATF3, DNAJB4, GCLM, HSPA6 and HSPH1) related to cellular stress response were significantly up-regulated in RhE model after 6 h treatment with representative skin sensitizers, 1-fluoro-2,4-dinitrobenzene and oxazolone, but not a non-sensitizer, benzalkonium chloride. The predictive performance of five genes was examined with eight skin sensitizers (e.g., cinnamic aldehyde), four non-sensitizers (e.g., sodium lauryl sulfate) and four pre-/pro-haptens (e.g., p-phenylenediamine, isoeugenol). When the positive criteria were set to obtain the highest accuracy with the animal testing (LLNA), ATF3, DNAJB4 and GCLM exhibited a high predictive accuracy (100%, 93.8% and 87.5%, respectively). All tested pre-/pro-haptens were correctly predicted by both ATF3 and DNAJB4. These results suggested that the RhE-based assay, termed epidermal sensitization assay (EpiSensA), could be an useful skin sensitization assay with a broad applicability domain including pre-/pro-haptens.