High-performance liquid chromatography LTQ-Orbitrap mass spectrometry method for tomatidine and non-target metabolites quantification in organic and normal tomatoes

Tomatoes, members of the Solanaceae plant family, produce biologically active secondary metabolites, including glycoalkaloids and aglycons, which may have both adverse and beneficial biological effects. A new liquid chromatography method that utilized LTQ-Orbitrap MS was developed for the analysis of tomatidine, the main aglycon in tomatoes. Recoveries of tomatidine were >98.3% with the relative standard deviations (RSDs) below 6.1%. The limit of detection (LODs) was 0.0003?mg?kg^?1. The limit of quantitation (LOQs) is 0.001?mg?kg^?1. The linear range was between with 0.0025 and 1?mg?kg^?1 with an excellent correlation coefficient (R²) equal to 0.9990. Various tomato samples were analyzed and the level of tomatidine in the 11 samples analysed was higher in normal respect to organic tomatoes. The capability of the set-up Full Scan LTQ-Orbitrap MS method allowed us to quantified two non-target analytes. The m/z 1032 was identified as dehydrotomatine, confirmed through accurate mass studies (mass error in ppm equal to 1.5017) meanwhile m/z 902 as (Glc)₂–Gal-Tomatidine (β₁-Tomatine) (mass error in ppm equal to 2.0719).     

Development of an ELISA for detection of Sudan I in food samples using monoclonal antibody

A highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed using a new monoclonal antibody for detecting the food colourant Sudan I. The half-maximum inhibition concentrations (IC₅₀) and the limit of detection (calculated as IC₂₀) of ELISA for Sudan I were 2 and 0.01 ng mL^?1, respectively. The study showed little cross-activity with Sudan I structural analogues (below 0.01%). The average recoveries in intra- and interassays for Sudan I from fortified fresh tomato and chilli samples by ELISA were in ranges of 82–94% and 79–91%, respectively. The coefficients of variation of intra- and interassays were 6–8% and 6–10%, respectively. The IC₅₀ was approximately 2.0 ng mL^?1 after the Sudan I-Kit had been kept for 180 days.     

外周血流式细胞仪分析在淋巴瘤性红皮病诊断中的作用研究

【摘要】 目的 探讨外周血流式细胞仪检测在红皮病诊断中的应用价值。方法 收集2017年9月至2020年12月在中国医学科学院皮肤病医院就诊的29例红皮病患者,包括6例红皮病型蕈样肉芽肿（EMF）、5例Sézary综合征（SS）及18例不同病因的炎症性红皮病（IE）,4例健康志愿者为健康对照。采用流式细胞仪检测外周血淋巴细胞亚群、免疫表型及克隆性,比较其在炎症性红皮病与淋巴瘤相关红皮病的差异。采用单因素方差分析及LSD-t检验进行组间比较。结果 4组受试者的T细胞、B细胞、NK细胞及CD4-CD8-细胞比例差异均有统计学意义（均P < 0.001）,SS患者T细胞比例（93.8% ± 3.4%）高于EMF（42.7% ± 6.4%）及IE（46.0% ± 6.8%,t = 12.8、14.4, P < 0.001）,IE患者CD4-CD8-细胞比例（0.37% ± 0.40%）低于EMF（2.93% ± 0.84%）及SS（2.38% ± 0.74%,t = 9.2、6.7, P < 0.05）。健康对照及IE患者均未检测到克隆性T细胞受体可变区β链（TCR-vβ）表达;3例 EMF及所有SS患者检测到表达克隆性TCR-vβ的细胞亚群,且TCR-vβ克隆性细胞均为CD4+CD7-CD26-表型。4组受试者CD4+ T淋巴细胞表面表达趋化因子受体CCR4、CXCR3、CCR5与皮肤淋巴细胞抗原（CLA）及程序性死亡受体1（PD-1）的细胞比例差异均有统计学意义（均P < 0.001）,IE患者表达CCR4、CLA、PD-1细胞比例低于SS及EMF患者（均P < 0.001）,表达CXCR3、CCR5细胞比例高于SS患者及EMF患者 （均P < 0.001）。 结论 流式细胞仪检测外周血淋巴细胞亚群、免疫表型及克隆性,可为红皮病患者的病因诊断提供佐证,有助于淋巴瘤相关红皮病与炎症性红皮病的鉴别诊断。     

Removal of biofilm is essential for long-term ventilation tube retention

BACKGROUND Although long-term retention of a ventilation tube is required in many ear diseases,spontaneous removal of conventional ventilation tube is observed in patients within 3 to 12 mo.To address this issue,we aimed to determine a new method for long-term retention of the ventilation tube.AIM To explore the value of removing the biofilm for long-term retention of tympanostomy ventilation tubes.METHODS A case-control study design was used to evaluate the safety and effectiveness of long-term tube retention by directly removing the biofilm(via surgical exfoliation)in patients who underwent myringotomy with ventilation tube placement.The patients were randomly divided into two groups:Control group and treatment group.Patients in the treatment group underwent regular biofilm exfoliation surgery in the clinic,whereas those in the control group did not have their biofilm removed.Only conventional ventilation tubes were placed in this study.Outcome measures were tube position and patency.Tube retention time and any complications were documented.RESULTS Eight patients with biofilm removal and eight patients without biofilm removal as a control group were enrolled in the study.The tympanostomy tube retention time was significantly longer in the treatment group(43.5±26.4 mo)than in the control group(9.5±6.9 mo)(P=0.003).More tympanostomy tubes were found to be patent and in correct position in the treatment group during the follow-up intervals than in the control group(P=0.01).CONCLUSION Despite the use of short-term ventilation tubes,direct biofilm removal can be a well-tolerated and effective treatment for long-term tube retention of tympanostomy ventilation tubes in patients who underwent myringotomy.     

肿瘤坏死因子β基因多态性与吉兰-巴雷综合征不同亚型的关联研究

目的研究肿瘤坏死因子βTNFβ基因多态性与吉兰-巴雷综合征(GBS)的两种主要亚型-急性炎症性脱髓鞘性多发性神经病(AIDP)和急性运动性轴索型神经病(AMAN)的易感性和预后的关系。方法对我院在2008至2011年收治的吉兰-巴雷综合征患者进行电生理标准分型,在疾病高峰期和发病1年后随访时进行Hughes神经功能评分;采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术对41例AIDP患者、52例AMAN患者和98名健康人进行TNFβ基因多态性分析。结果 AIDP组TNFβ基因型分布与对照组没有差别,TNFβG等位基因频率明显高于对照组(37.8%,25%);AMAN组TNFβ基因型分布与对照组相比有差别,TNFβG等位基因频率明显高于对照组(40.4%,25%),P<0.05;GBS重症型患者(Hughes评分4⁶分)TNFβG等位基因频率明显高于对照组(40.6%,25%),P<0.05。GBS预后较差患者(Hughes评分>2分)TNFβG等位基因频率明显高于对照组(47.5%,25%),P<0.05。结论 TNFβG等位基因与GBS两种亚型的易患性有关,携带有TNFβG等位基因患者更倾向有较严重的疾病病程且预后较差。     

Association of an Alphasatellite with Tomato Yellow Leaf Curl Virus and Ageratum Yellow Vein Virus in Japan is Suggestive of a Recent Introduction

Samples were collected in 2011 from tomato plants exhibiting typical tomato leaf curl disease symptoms in the vicinity of Komae, Japan. PCR mediated amplification, cloning and sequencing of all begomovirus components from two plants from different fields showed the plants to be infected by Tomato yellow leaf curl virus (TYLCV) and Ageratum yellow vein virus (AYVV). Both viruses have previously been shown to be present in Japan, although this is the first identification of AYVV on mainland Japan; the virus previously having been shown to be present on the Okinawa Islands. The plant harboring AYVV was also shown to contain the betasatellite Tomato leaf curl Java betasatellite (ToLCJaB), a satellite not previously shown to be present in Japan. No betasatellite was associated with the TYLCV infected tomato plants analyzed here, consistent with earlier findings for this virus in Japan. Surprisingly both plants were also found to harbor an alphasatellite; no alphasatellites having previously been reported from Japan. The alphasatellite associated with both viruses was shown to be Sida yellow vein China alphasatellite which has previously only been identified in the Yunnan Province of China and Nepal. The results suggest that further begomoviruses, and their associated satellites, are being introduced to Japan. The significance of these findings is discussed.     

乙型肝炎病毒颗粒性抗原转基因番茄植株的构建及鉴定

目的 构建乙型肝炎病毒(HBV)颗粒性抗原转基因番茄植株。方法 将乙型肝炎病毒preS／S基因插入到HBc的第73～94aa处(棘突尖部)，再把整个复合基因克隆到植物表达载体pBIN438(含35s起动子)中，通过液氮冻融法转化根瘤农杆菌ehal05，采用叶盘法转化番茄。结果 得到一批转基因番茄植株，经PCR、PCR-Southern blot和Souhtem blot证实HBc-HBs基因已整合到番茄基因组中；经Western blot证实HBc-HBs能在番茄中有效表达。结论 成功构建了乙肝颗粒性抗原基因的转基因番茄植株，为下一步进行该番茄疫苗株的效果评价奠定了基础。     

Consumption of Fruitflow® lowers blood pressure in pre-hypertensive males: a randomised,placebo controlled,double blind,cross-over study

In order to investigate whether the angiotensin converting enzyme-inhibitory tomato extract Fruitflow^® would lower blood pressure after consumption, we conducted a randomised, double-blinded, placebo-controlled human intervention study, involving 12 pre-hypertensive people in a crossover design. Consuming a single dose of 150?mg Fruitflow^® resulted in a significant reduction in 24-hour average blood pressure as well as average wake-period and sleep-period SBP, compared to placebo. Other parameters related to blood pressure, such as 24-hour average mean arterial pressure, pulse pressure, heart rate, central aortic systolic pressure and radial augmentation index were also reduced. In addition, the platelet aggregation response to ADP, measured 24?hours after consuming Fruitflow^®, fell significantly compared to baseline, and compared to placebo. This pilot study clearly shows the beneficial effects of Fruitflow^® on two important cardiovascular risk factors, high blood pressure and platelet hyperactivity.     

泽泻汤组分不同配比对豚鼠膜迷路积水影响的比较

目的:观察泽泻汤中泽泻与白术不同剂量配比对膜迷路积水豚鼠的影响。方法:采用腹腔注射醋酸去氨加压素法复制膜迷路积水动物模型。观察各组豚鼠的行为学变化及蜗管组织病理学改变,计算各组豚鼠蜗管横截面积与蜗管加前庭面积之和(总面积)的比值,以评价膜迷路积水的程度。结果:与空白组比较,模型组豚鼠蜗管横截面积与总面积的比值明显升高(P<0.05);与模型组比较,3组实验组豚鼠蜗管横截面积与总面积的比值均下降,但实验1组与模型组比较差异有统计学意义(P<0.05)。结论:3组实验组中,以原方剂量配比(泽泻∶白术为5∶2)的实验1组缓解豚鼠膜迷路积水作用最佳,证实了仲景方剂配伍的科学合理性。