3,5-O-二咖啡酰基奎宁酸的制备及其对HeLa细胞的抗增殖活性研究

目的 制备高纯度3，5-O-二咖啡酰基奎宁酸，并评价其对人宫颈癌HeLa细胞的抗增殖活性。方法 采用柱色谱提取法和中压液相色谱法从奇蒿花中分离、纯化得到高纯度的3，5-O-二咖啡酰基奎宁酸。采用MTT法评价该化合物对人宫颈癌HeLa细胞的体外抗增殖活性。结果 柱色谱提取的提取率和中压液相色谱法的回收率分别为99.0%，61.2%，总回收率为54.0%。随着3，5-O-二咖啡酰基奎宁酸浓度升高，HeLa细胞存活率下降，细胞形态损伤增加，受试药物的IC₅₀值为26.5µg·mL^-1。结论 本研究提供了一种简单、高效、节能的3，5-O-二咖啡酰基奎宁酸制备方法。3，5-O-二咖啡酰基奎宁酸对HeLa细胞具有一定的体外抗增殖活性。     

基于德尔菲法构建呼吸与危重症医学科住院医师规范化培训出科考核指标体系

目的　根据临床需求，结合医院实际，建立一套适应呼吸与重症医学科发展的科学、系统的出科考核评价体系。方法　采用文献分析法和德尔菲法，经过两轮咨询确定指标体系，采用优序图法确定指标权重。采用SPSS 22.0统计软件进行数据分析。结果　两轮咨询专家积极系数为100%和95.65%；两次咨询群体专家权威系数平均值为0.85；两轮专家咨询的协调系数分别为0.513、0.516；最终确立一级指标5项、二级指标14项。结论　专家积极程度和协调系数好，结果可信。确立的指标体系可用于医院呼吸与危重症医学科住院医师规范化培训考核评估。     

Mobile Intensive Care Unit versus Hospital walk-in patients,in the treatment of first episode ST- elevation myocardial infarction

ObjectiveTo determine the impact of the attention given by emergency medical services teams working in mobile intensive care units (MICU) versus patients arriving at the hospital under their own means with ST-elevation myocardial infarction (STEMI) event in terms of time to reperfusion (TR), mortality at 30 days and six months.MethodsWe retrospectively studied 634 consecutive patients with STEMI who underwent primary a percutaneous coronary intervention from January 1st 2015 to December 31st 2018 in a single centre. Depending on the first medical contact patients were classified into two groups, MICU versus walk-in patients. We extracted data on patients’ characteristics, symptoms, treatments, times to reperfusion and mortality.ResultsIn our study 634 patients were included, of whom 59.0% were initially attended by the MICU. Differences were seen between the two groups in time delays to the first medical contact (120.0 vs 63.0 min; p < 0.001) and TR (208.0 Vs 150.0 min; p < 0.001). Patients attended by the MICUs presented a shorter ICU and hospital stay. The lowest 30-day mortality rate was observed in MICU group: 9.0% in contrast with 4.5%, p = 0.03; remaining after 6 months. The multivariable analysis showed that the initial attention given by MICU to STEMI patients was a protective agent against mortality [OR: 0.32 (0.11–0.90); p = 0.03].ConclusionInitial attention of the patients with STEMI by doctor-on-board-MICU and available 24 h a day 7 days a week as part of a regional network (CORECAM), was associated with a decrease in the ischemia time, hospital stay and mortality of these patients in our environment.     

UVB下调IFN-γ诱导的正常人角质形成细胞表面抗原的表达

目的：研究中波紫外线（UVB）对IFN-γ体外诱导的正常人角质形成细胞(KC)表面抗原表达的影响及可能机制。方法：分离培养原代人KC，用适当浓度的IFN-γ对细胞进行预处理，再用生理剂量的UVB照射细胞。流式细胞术测定照射前后细胞表面CD40、CD54、MHC-II的表达，放射免疫法测定照射前后细胞上清液中α-MSH浓度，ELISA测定上清液中IL-6、IL-8、TNF-α等细胞因子浓度。结果：KC低水平表达CD40、CD54、MHC-II抗原，IFN-γ处理后，细胞活性增强，表面抗原的表达显著增加，再用UVB照射后，细胞活性下降，表面抗原表达明显下调，而上清液中α-MSH的浓度则逐渐上升，照后72 h达最高峰。KC经可溶性CD40配体(sCD40L)处理后，IL-8的释放和CD54的表达加强。结论：UVB能抑制IFN-γ诱导的KC表面抗原的表达，这可能与UVB诱导KC自分泌产生α-MSH等免疫抑制物有关。     

Berberine diminishes cancer cell PD-L1 expression and facilitates antitumor immunity via inhibiting the deubiquitination activity of CSN5

Programmed cell death-1 (PD-1)/programmed cell death ligand-1 (PD-L1) blocking therapy has become a major pillar of cancer immunotherapy. Compared with antibodies targeting, small-molecule checkpoint inhibitors which have favorable pharmacokinetics are urgently needed. Here we identified berberine (BBR), a proven anti-inflammation drug, as a negative regulator of PD-L1 from a set of traditional Chinese medicine (TCM) chemical monomers. BBR enhanced the sensitivity of tumour cells to co-cultured T-cells by decreasing the level of PD-L1 in cancer cells. In addition, BBR exerted its antitumor effect in Lewis tumor xenograft mice through enhancing tumor-infiltrating T-cell immunity and attenuating the activation of immunosuppressive myeloid-derived suppressor cells (MDSCs) and regulatory T-cells (Tregs). BBR triggered PD-L1 degradation through ubiquitin (Ub)/proteasome-dependent pathway. Remarkably, BBR selectively bound to the glutamic acid 76 of constitutive photomorphogenic-9 signalosome 5 (CSN5) and inhibited PD-1/PD-L1 axis through its deubiquitination activity, resulting in ubiquitination and degradation of PD-L1. Our data reveals a previously unrecognized antitumor mechanism of BBR, suggesting BBR is small-molecule immune checkpoint inhibitor for cancer treatment.     

甲状腺疾病患者血清 Tg-CIC 中 Tg-Ab 水平检测及相关临床研究

目的　对血清甲状腺球蛋白免疫复合物 (thyroglobulin circulating immune complex, Tg-CIC) 中抗甲状腺球蛋白抗体 (anti-thyroglobulin antibody, Tg-Ab) 含量检测，探讨 Tg-CIC 在甲状腺疾病中的临床以及流行病学意义。方法　收集甲状腺疾病患者血清标本 187 例，每例血清标本分为两组 ( 实验组和空白对照组 )，采用免疫复合物解离专利技术对甲状腺疾病患者血清标本 Tg-CIC 进行分离、解离，通过检测 Tg-CIC 解离后的 Tg-Ab 含量来间接反映甲状腺疾病患者血清中Tg-CIC 水平，按照不同的甲状腺疾病进行分组并对实验结果分析讨论。结果　不同甲状腺患者血清中 Tg-CIC 总阳性率达 92.51%，不同甲状腺疾病血清中 Tg-CIC 的阳性率差异无统计学意义 (χ?=2.917, P>0.05)。不同甲状腺疾病患者间血清游离 Tg-Ab 含量差异无统计学意义 (H=3.882, P>0.05)，不同甲状腺疾病患者间血清 CIC 中 Tg-Ab 含量差异无统计学意义(H=5.5842, P>0.05)。不同甲状腺疾病患者中，甲状腺功能亢进、甲状腺炎以及甲状腺结节患者血清游离 Tg-Ab 的含量与血清 CIC 中的 Tg-Ab 含量呈正相关 (P<0.05)，甲状腺功能减退、甲状腺肿瘤患者血清游离 Tg-Ab 的含量与血清 CIC 中的Tg-Ab 含量不相关 (P>0.05)。结论　在甲状腺疾病患者血清中，大部分可检出 Tg-CIC，与疾病类型无关。而甲状腺疾病患者中，甲状腺功能亢进、甲状腺炎以及甲状腺结节患者血清游离 Tg-Ab 含量与血清 CIC 中 Tg-Ab 含量存在相关性，为我们进一步了解 Tg-CIC 与甲状腺疾病的发生、发展的相关性研究奠定了基础，提供了新的研究视角。