孟河医派特色炮制猪心血拌丹参及其他炮制品活性成分与抗脑缺血氧化损伤相关性分析

目的 分析猪心血拌丹参及其他丹参炮制品中11个小分子活性成分及1个蛋白成分与抗脑缺血氧化损伤的相关性,明确猪心血拌丹参抗脑缺血氧化损伤的关键成分标志物。方法 采用高效液相色谱法（HPLC）同时测定丹参、猪心血拌丹参、猪血拌丹参、酒拌丹参、猪血清转铁蛋白（Tf）拌丹参5种饮片中11种活性成分含量;采用酶联免疫吸附测定法（ELISA）检测丹参不同炮制品中Tf的含量。构建斑马鱼缺血性脑卒中模型,考察丹参不同炮制品对脑缺血斑马鱼行为轨迹、转基因斑马鱼Tg（elavl3：eGFP）脑神经元损伤及脑组织中丙二醛（MDA）和超氧化物歧化酶（SOD）水平的影响;构建糖氧剥夺/复氧（OGD/R）诱导的小鼠海马神经细胞缺血缺氧模型,以乳酸脱氢酶（LDH）、活性氧（ROS）、MDA和SOD为评价指标,考察丹参不同炮制品对神经细胞氧化损伤的影响;通过主成分分析（PCA）、偏最小二乘判别法-判别分析（PLS-DA）及Spearman相关性分析对11个小分子活性成分及1个蛋白成分与药效指标进行分析,筛选猪心血拌丹参抗脑缺血氧化损伤的关键成分标志物。结果 与丹参比较,丹参各炮制品中水溶性、脂溶性成分含量均呈不同程度的升高,而丹酚酸A成分含量降低;与酒拌丹参比较,猪心血拌丹参、猪血拌丹参及Tf拌丹参中丹酚酸B、丹参素、迷迭香酸等成分含量均升高,而丹酚酸A含量降低。ELISA结果显示,猪心血拌丹参、猪血拌丹参及Tf拌丹参中均含有Tf蛋白,且三者含量差异无统计学意义。药效结果显示,丹参不同炮制品均能改善斑马鱼缺血性脑卒中后出现的行为缺陷、脑神经元损伤及氧化应激,且猪心血拌丹参组作用最明显。PCA结果显示,丹酚酸B、丹酚酸A、迷迭香酸、紫草酸、丹参素、咖啡酸、丹参酮Ⅱ_A、隐丹参酮、丹参酮Ⅰ为丹参及其炮制品中的主要贡献成分。相关性分析结果显示,隐丹参酮、迷迭香酸、咖啡酸、二氢丹参酮Ⅰ、丹酚酸B、丹参酮Ⅱ_A、丹参酮Ⅰ与斑马鱼脑组织中MDA水平呈负相关,紫草酸、原儿茶醛、迷迭香酸、二氢丹参酮Ⅰ、丹酚酸B、Tf与SOD水平呈正相关,迷迭香酸、咖啡酸、二氢丹参酮Ⅰ、丹酚酸B、丹参酮Ⅱ_A、丹参酮Ⅰ、丹参素、Tf与斑马鱼脑神经元荧光强度呈正相关;紫草酸、原儿茶醛、迷迭香酸、二氢丹参酮Ⅰ、丹酚酸B、丹参素、Tf与HT22细胞中LDH、ROS、MDA水平呈负相关,与SOD水平呈正相关。结论 丹参及其不同炮制品抗脑缺血氧化损伤作用存在差异,其中以猪心血拌丹参改善氧化损伤作用最强,这可能与丹酚酸B、丹参素、迷迭香酸等成分的含量变化有关,可为明确该孟河医派特色炮制品抗脑缺血质量标志物及阐明其炮制机制提供依据。     

Pre-attack and pre-episode symptoms in cluster headache: a multicenter cross-sectional study of 327 Chinese patients

BackgroundThere have been a few studies regarding the pre-attack symptoms (PAS) and pre-episode symptoms (PES) of cluster headache (CH), but none have been conducted in the Chinese population. The purpose of this study was to identify the prevalence and features of PAS and PES in Chinese patients, as well as to investigate their relationships with pertinent factors.MethodsThe study included patients who visited a tertiary headache center and nine other headache clinics between January 2019 and September 2021. A questionnaire was used to collect general data and information about PAS and PES.ResultsAmong the 327 patients who met the CH criteria (International Classification of Headache Disorders, 3rd edition), 269 (82.3%) patients experienced at least one PAS. The most common PAS were head and facial discomfort (74.4%). Multivariable logistic regression analysis depicted that the number of triggers (OR = 1.798, p = 0.001), and smoking history (OR = 2.067, p = 0.026) were correlated with increased odds of PAS. In total, 68 (20.8%) patients had PES. The most common symptoms were head and facial discomfort (23, 33.8%). Multivariable logistic regression analysis showed that the number of triggers were associated with increased odds of PES (OR = 1.372, p = 0.005).ConclusionsPAS are quite common in CH patients, demonstrating that CH attacks are not comprised of a pain phase alone; investigations of PAS and PES could help researchers better understand the pathophysiology of CH.     

The cell cycle inhibitor P21 promotes the development of pulmonary fibrosis by suppressing lung alveolar regeneration

The cell cycle inhibitor P21 has been implicated in cell senescence and plays an important role in the injury–repair process following lung injury. Pulmonary fibrosis (PF) is a fibrotic lung disorder characterized by cell senescence in lung alveolar epithelial cells. In this study, we report that P21 expression was increased in alveolar epithelial type 2 cells (AEC2s) in a time-dependent manner following multiple bleomycin-induced PF. Repeated injury of AEC2s resulted in telomere shortening and triggered P21-dependent cell senescence. AEC2s with elevated expression of P21 lost their self-renewal and differentiation abilities. In particular, elevated P21 not only induced cell cycle arrest in AEC2s but also bound to P300 and β-catenin and inhibited AEC2 differentiation by disturbing the P300–β-catenin interaction. Meanwhile, senescent AEC2s triggered myofibroblast activation by releasing profibrotic cytokines. Knockdown of P21 restored AEC2-mediated lung alveolar regeneration in mice with chronic PF. The results of our study reveal a mechanism of P21-mediated lung regeneration failure during PF development, which suggests a potential strategy for the treatment of fibrotic lung diseases.     

Emerging pro-drug and nano-drug strategies for gemcitabine-based cancer therapy

Gemcitabine has been extensively applied in treating various solid tumors. Nonetheless,the clinical performance of gemcitabine is severely restricted by its unsatisfactory pharmacokinetic parameters and easy deactivation mainly because of its rapid deamination, deficiencies in deoxycytidine kinase (DCK), and alterations in nucleoside transporter. On this account, repeated injections with a high concentration of gemcitabine are adopted, leading to severe systemic toxicity to healthy cells. Accord...     

Identification of DAPI-stained normal,inflammatory, and carcinoma hepatic cells based on hyperspectral microscopy

Gross chromatin imbalance and high DNA content are distinct features of various types of cancer cells. However, severe inflammation can also produce similar symptoms in cells. In this study, normal, inflammatory, and carcinoma hepatic cells were stained with 4’,6-diamidino-2-phenylindole (DAPI) and investigated by hyperspectral microscopy. DAPI is a DNA-sensitive fluorochrome. Therefore, the differences in the cellular DNA of the samples can be revealed by the corresponding fluorescence. Our experimental results demonstrate that although chromosomal disorder and high DNA content both occur in severely inflammatory and carcinoma hepatic cells, there is still a slight difference in their DNA, making their fluorescent intensity and even their spectral shapes distinguishable. Based on these spectral features, we developed a method for the precise identification of normal, inflammatory, and carcinoma hepatic cells in the field of view. The identification accuracy for these three types of cells was 99.8%. We believe that examination that combines DAPI staining with hyperspectral microscopy is a potential method for the identification and investigation of various types of cancer tissues.     

30 d-6°头低位卧床对人下肢脂肪体积的影响

目的 观察模拟失重对人下肢脂肪体积的影响,验证磁共振水脂分离成像技术在测量人下肢脂肪容积中的可行性.方法 14名男性健康志愿者,-6°头低位卧床30 d,使用磁共振水脂分离成像(Dixon)技术采集卧床前后的下肢横断面解剖图像,测量卧床前后下肢脂肪体积.结果 与卧床前相比,卧床后所有志愿者下肢总脂肪体积及大腿段脂肪体积显著增加(P＜0.001);小腿段脂肪体积呈增加趋势,但无统计学差异(P＞0.05).结论 使用MRI水脂分离成像可以清晰地显示卧床前后下肢脂肪体积的变化;-6°头低位卧床30 d后下肢脂肪体积明显增加.     

彩色多普勒超声对医源性股动脉假性动脉瘤的临床应用

目的：探讨彩色多普勒超声对医源性股动脉假性动脉瘤的诊断及治疗价值。方法对22例经皮股动脉穿刺介入术后触及搏动性肿块的患者采用二维超声、彩色多普勒超声、频谱多普勒超声检查,对声像图进行总结分析,在超声引导下压迫及注射治疗。结果本组22例假性动脉瘤彩色多普勒超声诊断与临床符合率100%（22/22）,18例经超声引导下压迫治疗后瘤腔闭合,3例经瘤腔内注射凝血酶治疗取得满意疗效,总治愈率95%（21/22）,1例行外科手术治疗。结论彩色多普勒超声是医源性假性动脉瘤首选的诊断及治疗方法。