Investigation of cell cytotoxic activity and molecular mechanism of 5β,19-epoxycucurbita-6,23(E)-diene-3β,19(R),25-triol isolated from Momordica charantia on hepatoma cells

ContextMomordica charantia L. (Cucurbitaceae), known as bitter melon, is an edible fruit cultivated in the tropics. In this study, an active compound, 5β,19-epoxycucurbita-6,23(E)-diene-3β,19(R),25-triol (ECDT), isolated from M. charantia was investigated in regard to its cytotoxic effect on human hepatocellular carcinoma (HCC) cells.ObjectiveTo examine the mechanisms of ECDT-induced apoptosis in HCC cells.Materials and methodsThe inhibitive activity of ECDT on HA22T HCC cells was examined by MTT assay, colony formation assay, wound healing assay, TUNEL/DAPI staining, annexin V-fluorescein isothiocyanate/propidium iodide (PI) staining and JC-1 dye. HA22T cells were treated with ECDT (5, 10, 15, 20 and 25 μM) for 24 h, and the molecular mechanism of cells apoptosis was examined by Western blot. Cells treated with vehicle DMSO were used as the negative control.ResultsECDT inhibited the cell proliferation of HA22T cells in a dose-dependent manner. Flow cytometry showed that ECDT treatment at 10–20 μM increased early apoptosis by 10–14% and late apoptosis by 2–5%. Western blot revealed that ECDT treatment activated the mitochondrial-dependent apoptotic pathway, and ECDT-induced apoptosis was mediated by the caspase signalling pathway and activation of JNK and p38MAPK. Pre-treatment of cells with MAPK inhibitors (SB203580 or SP600125) reversed the ECDT-induced cell death, which further supported the involvement of the p38MAPK and JNK pathways.Discussion and conclusionsOur results indicated that ECDT can induce apoptosis through the p38MAPK and JNK pathways in HA22T cells. The findings suggested that ECDT has a valuable anticancer property with the potential to be developed as a new chemotherapeutic agent for the treatment of HCC.     

Precise immunological evaluation rationalizes the design of a self-adjuvanting vaccine composed of glycan antigen,TLR1/2 ligand,and T-helper cell epitope

Sialyl-Tn (STn), overexpressed on various tumors, has been investigated for its application in anti-cancer vaccine therapy. However, Theratope, an STn-based vaccine, failed in the phase III clinical trial due to poor immunogenicity and epitope suppression by the foreign carrier protein. We therefore developed a self-adjuvanting STn based-vaccine, a conjugate of clustered STn (triSTn) antigen, TLR1/2 ligand (Pam₃CSK₄), and T-helper (Th) cell epitope, and found that this three-component self-adjuvanting vaccine effectively resulted in the production of anti-triSTn IgG antibodies. We herein analyzed immune responses induced by this self-adjuvanting vaccine in detail. We newly synthesized two-component vaccines, i.e., Pam₃CSK₄- or Th epitope-conjugated triSTn, as references to evaluate the immune-stimulating functions of Pam₃CSK₄ and Th epitope. Immunological evaluation of the synthesized vaccine candidates revealed that Pam₃CSK₄ was essential for antibody production, indicating that the uptake of triSTn antigen by antigen-presenting cells (APCs) was promoted by the recognition of Pam₃CSK₄ by TLR1/2. The function of the Th epitope was also confirmed. Th cell activation was important for boosting antibody production and IgG subclass switching. Furthermore, flow cytometric analyses of immune cells, including T cells, B cells, dendritic cells, and other monocytes, were first employed in the evaluation of self-adjuvanting vaccines and revealed that the three-component vaccine was able to induce antigen-specific immune responses for efficient antibody production without excessive inflammatory responses. Importantly, the co-administration of Freund''s adjuvants was suggested to cause excessive myeloid cell accumulation and decreased plasma cell differentiation. These results demonstrate that vaccines can be designed to achieve the desired immune responses via the bottom-up construction of each immune element.

Detailed analysis of a three-component self-adjuvanting vaccine revealed that conjugate vaccines can be designed to achieve the desired immune responses via bottom-up construction of the necessary immune elements.     

北京市郊区农村基层医疗卫生服务能力现状及问题分析

目的分析北京市郊区村巡诊和村卫生室建设现状,为完善农村基层医疗卫生服务体系建设提供参考。方法对北京市郊7个区中119个行政村进行实地观察,并对652名相关研究对象进行半结构化访谈,包括对巡诊和卫生室的满意度、基层医疗机构的服务质量等。结果村巡诊服务效率不高,不能满足居民实际医疗服务需求;村卫生室医疗人员短缺,基本药物配备不全,报销途径不畅通,村民就医行为更倾向于选择乡镇卫生院和县医院。结论为更好落实卫生领域重要民生项目,切实提升农村居民就医体验,应进一步落实和丰富村巡诊内容,加强基层全科医生培养力度,提升卫生费用投资的针对性和有效性,把村卫生健康服务情况纳入村干部工作考核指标,优化基本药物目录,扩大新农合定点报销覆盖范围等。     

1996～2006年我院抗癫痫药的用药状况分析

目的了解我院1996～2006年抗癫痫药的应用状况及变化趋势。方法采用WHO推荐的以DDD数为指标的药物利用研究方法,计算我院抗癫痫药的销售金额、用药频度、日用金额等指标并进行排序。结果我院抗癫痫药的销售金额和DDDs值大体上呈逐渐增长趋势。并且以传统抗癫痫药物为主,丙戊酸盐、卡马西平、苯妥英钠三种传统的抗癫痫药物临床使用频度约占95%。结论我们应该对药物的使用情况进行定期分析比较,为合理用药、开发新型抗癫痫药提供科学依据和思路。     

高压氧辅助治疗对乳腺癌术后皮瓣愈合及血管内皮生长因子和缺氧诱导因子的影响

目的:探究高压氧辅助治疗对乳腺癌手术后皮瓣愈合及血管内皮生长因子(VEGF)和缺氧诱导因子-1α(HIF-1α)的影响。方法:选取长治医学院附属和平医院乳腺科2017年1月至2019年6月收治的132例乳腺癌患者作为研究对象,按随机数字表法分为对照组和观察组(每组66例)。对照组采用乳腺癌改良根治术,观察组在对照组治疗...     

药剂学GMP车间实训线上教学的探索

药剂学GMP虚拟仿真实训教学已被应用于药剂学实训教学中。在新形势下,基于教育部倡议“停课不停学”的基本思想,如何实现教师和学生在校园外或家中也能完成药剂学GMP虚拟仿真实训教学和学习任务是虚拟仿真教学改革噬待解决的重要问题。线上教学为该问题的解决提出了新的思路,线上教学目前已经成为了各大学开展课程的重要技术手段。基于GMP虚拟仿真实训教学是基于虚拟仿真平台的一门课程,虚拟仿真技术平台可以网络共享的特点决定了其与线上教学结合的可能性。本文以虚拟仿真技术平台与线上教学联合应用为目标建设了药剂学GMP车间实训线上课程。首先建设以线上平台（钉钉）与网络版GMP车间实训虚拟仿真技术平台联合模式为框架的在线课堂,为教师和学生提供可供课堂交流的平台。有效的解决了学生与教师远程交流的问题,实现了虚拟仿真平台互动教学的目标。然后在线课堂发布课程资源以及完成GMP车间实训在线教学和学习任务。在钉钉平台开通直播教学,通过在线演示或播放视频的形式进行知识讲解和师生互动,让学生完成对实训教学内容的学习。文章设计的在线教学与虚拟仿真教学联合教学模式的尝试拓展了在线教学在高校虚拟仿真课程中的应用,也为虚拟仿真平台与...     

不同频率神经肌肉电刺激对ARDS相关性ICU-AW小鼠肌肉萎缩防治及临床意义

目的 不同频率下神经肌肉电刺激(neuromuscular electrical stimulation,NMES)在ARDS相关性ICU获得性衰弱(ICU-acquired weakness,ICU-AW)中的作用及机制。方法 健康雄性C57BL/6小鼠88只随机分为11组,每组8只。分为:空白对照组(C1)、气管内注入无菌水组(C2)、ICU-AW模型组(ICU-AW)、ICU-AW+AMPK激动剂A-769662组(ICU-AW-A)、ICU-AW+A-769662溶剂对照组(ICU-AWV)、NMES 20 Hz组(ICU-AW-20)、NMES 40 Hz组(ICU-AW-40)、NMES 60 Hz组(ICU-AW-60)、NMES80 Hz组(ICU-AW-80)、ICU-AW-40+AMPK抑制剂Compound C组(ICU-AW-40-C)、ICU-AW-40+Compound C溶剂对照组(ICU-AW-40-V)。检测小鼠四肢抓力和存活状态,7 d后收集小鼠肺组织和腓肠肌标本,采用HE染色观察肺和肌肉病理学变化,采用western blot以及qRT-PCR的方...     

迭代重建技术对双低剂量肺动脉CT血管造影图像质量分析

目的 分析迭代重建技术对双低剂量肺动脉CT血管造影(CTA)检查图像质量的影响。方法 选取2018年2月至2021年2月我院行肺动脉CTA检查73例,随机分为对照组35例,观察组38例。对照组采用常规剂量滤波反投影重建,观察组采用双低剂量iDose⁴迭代重建技术。比较两组图像参数[背景噪声值(BN)、肺动脉CT值平均值(SI_vessel)、信噪声比(SNR)、对比噪声比(CNR)]、辐射剂量[容积CT剂量指数(CTDI_vol)、剂量长度指数(DLP)、有效吸收剂量(ED)]及图像质量,并分析典型病例的影像学表现。结果 观察组BN、SI_vessel、SNR、CNR参数高于对照组(P<0.05);观察组CTDI_vol、DLP、ED辐射剂量低于对照组(P<0.05);观察组图像质量优良率94.74%高于对照组77.14%(P<0.05)。结论 迭代重建技术应用于双低剂量肺动脉CTA检查中能降低辐射剂量,提高信噪比,改善图像质量。