基于网络药理和细胞实验探索芪玉三龙方治疗非小细胞肺癌的机制

目的 采用网络药理学和细胞实验探究芪玉三龙方治疗非小细胞肺癌的潜在作用机制。方法 利用中药系统药理学数据库与分析平台和中医药整合数据库检索芪玉三龙方成分筛选靶点,通过美国肿瘤基因组图谱数据库获得肺癌靶点基因。构建蛋白相互作用网络,并采用基因本体数据库和京都基因和基因组百科全书数据库进行富集分析。对核心靶蛋白与化合物进行分子对接分析,并以A549细胞进行实验验证。结果 芪玉三龙方对于非小细胞肺癌的潜在有效靶点162个,蛋白相互作用网络分析得到核心靶点基因20个,主要的生物学过程为对缺氧的反应、MAPK级联、炎症反应,主要的功能通路为钙信号通路、TNF信号通路、IL-17信号通路、细胞周期。分子对接结果表明,MMP9-槲皮素、PPARG-异鼠李素、PTGS2-刺芒柄花素、JUN-刺芒柄花素之间存在有效的结合亲和力。在A549细胞实验中,PCR和Western blot分析结果表明,芪玉三龙方含药血清能够促进PPAR-γ mRNA和蛋白表达（P＜0.05）,抑制c-JUN、COX-2、MMP-9 mRNA和蛋白表达（P＜0.05）。结论 芪玉三龙方通过影响A549肺癌细胞的凋亡、侵袭和肿瘤血管生成及炎症进程达到干预肺癌的作用。     

六味地黄汤通过树突状细胞Notch信号通路调节T细胞分化干预自身免疫性脑脊髓炎

目的 通过观察树突状细胞(dendritic cells,DCs)Notch信号通路对CD4+T细胞分化的影响,探究六味地黄汤干预实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyeli-tis,EAE)的机制.方法 采用主动免疫法诱导2D2小鼠EAE模型,采用磁珠分选收集...     

幽门螺杆菌既往感染胃癌前状态的中医证候学特点探究

目的 基于《京都胃炎分类》中的内镜表现探讨幽门螺杆菌（helicobacter pylori,Hp）既往感染状态胃癌前状态的中医证候学特点。方法 按照《京都胃炎分类》中的方法,收集内镜下表现符合Hp既往感染胃癌前状态特征的205例患者的中医四诊信息,采用因子分析和聚类分析,浅析其中医证候学特点。结果 经过因子分析提取出15个公因子,主要病位证素为肝、脾、胃,涉及的病性证素为气虚、阴虚、阳虚、火热、湿浊、血瘀、血热、痰浊、食积,经聚类得出肝胃气滞兼脾气亏虚证、脾胃湿热兼气阴两伤证、湿热瘀阻证、胃气壅滞证、肝胃郁热证、肝郁脾虚证共6个证型。结论 应用《京都胃炎分类》中的内镜观察方法,结合Hp既往感染病史及病理结果,所获得的Hp既往感染状态下胃癌前状态的中医证候学特点,能够为中医药诊治本病提供较为准确的辨证依据和思路。     

益气活血通络方通过抑制P2X7介导的脊髓小胶质细胞活化改善糖尿病大鼠神经病理性疼痛

目的 探讨益气活血通络方对糖尿病神经病理性疼痛(diabetic neuropathic pain,DNP)的影响及其作用机制.方法 高脂喂养大鼠4周后,35 mg/kg链脲佐菌素(streptozotocin,STZ)腹腔注射复制2型糖尿病大鼠模型,通过观测大鼠机械痛阈和热缩足反应时间作为判定DNP模型复制成功的标准...     

生血方拮抗奥沙利铂联合替吉奥方案致晚期胃癌骨髓抑制的临床观察

目的 观察自拟生血方治疗脾肾两虚型晚期胃癌行奥沙利铂联合替吉奥方案化学治疗致骨髓抑制的疗效和安全性。方法 将60例中医辨证为脾肾两虚型晚期胃癌患者随机分成对照组和观察组,每组30例。对照组予以奥沙利铂联合替吉奥方案化学治疗,观察组在对照组基础上加用自拟生血方汤剂口服及粉剂加工后外敷穴位联合治疗。观察两组患者首次骨髓抑制发生时间、骨髓抑制持续时间、0~3周骨髓抑制等级变化,采用卡氏功能状态量表（Karnofskys performance scale,KPS）、中医证候积分评价患者生活质量,观察两组患者的无进展生存期（progression- free survival, PFS）。结果 与对照组比较,观察组患者首次出现白细胞、血红蛋白、血小板下降的时间明显延长（P＜0.05）,白细胞、血红蛋白、血小板抑制持续时间明显缩短（P＜0.05）。两组患者不同时点白细胞、血红蛋白、血小板抑制等级比较,差异有统计学意义（P＜0.05）,两组患者化学治疗第21天与化学治疗第1天白细胞抑制等级变化比较,差异有统计学意义（P＜0.05）。与治疗前比较,对照组患者治疗后KPS评分显著降低（P＜0.05）。两组患者治疗前后KPS评分差值比较,差异有统计学意义（P＜0.05）。与治疗前比较,观察组患者中医证候积分显著下降（P＜0.05）,观察组中医证候积分下降程度明显大于对照组（P＜0.05）。观察组患者PFS为10个月,较对照组PFS（6个月）明显延长。结论 自拟生血方治疗晚期脾肾两虚型胃癌奥沙利铂联合替吉奥方案化学治疗所致骨髓抑制,能延迟首次骨髓抑制发生时间并缩短持续时间,降低白细胞骨髓抑制等级,在一定程度上改善晚期胃癌患者临床症状及生活质量。     

新藤黄酸抑制人肝癌细胞BEL-7402自噬作用研究

目的观察新藤黄酸(gambogenic acid,GNA)对人肝癌细胞BEL-7402自噬的影响,为GNA抗肿瘤研究提供理论依据。方法体外培养人肝癌BEL-7402细胞株,采用MTT法检测GNA对人肝癌细胞BEL-7402存活率的影响;倒置显微镜观察给药前后细胞形态变化;单丹磺酰尸胺(monodansylcadaverine,MDC)染色,荧光显微镜观察细胞自噬小体;透射电子显微镜进一步观察自噬小体;Western blot法检测自噬相关蛋白微管相关蛋白1轻链3(microtubule associated protein 1light chain 3,LC3)和p62、Beclin1的表达水平及加入自噬诱导剂雷帕霉素(rapamycin,Rap)和自噬抑制剂氯喹(chloroquine,CQ)后自噬相关蛋白的表达情况。结果 GNA能显著抑制人肝癌细胞BEL-7402的增殖并呈时效及量效关系;倒置显微镜观察结果显示GNA组细胞随药物浓度增大,细胞悬浮增多,高浓度给药组细胞出现碎片化;荧光显微镜下观察结果显示,随药物浓度的增加,荧光点状聚集增多;透射电子显微镜观察发现,随GNA浓度的增加,细胞内自噬空泡数量增多,与对照组比较,加入自噬诱导剂Rap后,自噬空泡数量显著增多;Western blot法检测结果显示,随GNA浓度的增加,Beclin1、p62、LC3-Ⅱ蛋白表达水平均增加(P<0.05),LC3-Ⅰ无明显变化,表明BEL-7402细胞自噬前期被激活,后期进程被抑制。结论 GNA能抑制人肝癌细胞BEL-7402的增殖,其机制可能部分是通过抑制自噬后期自噬体与溶酶体的融合,抑制人肝癌BEL-7402细胞的保护性自噬。     

HSP27慢病毒载体的构建及其稳定表达细胞株的建立

热休克蛋白27(HSP27)是一种结构上高度保守的小分子热休克应激蛋白,它可通过与细胞自噬、细胞凋亡和天然免疫信号通路关键因子相互结合参与多种病毒的感染过程.为了便于深入研究HSP27在机体抗感染免疫中的作用,用PCR方法从A549细胞中扩增出HSP27片段,然后经双酶切、连接、转化等步骤获得HSP27的慢病毒表达载体...     

Forensic Autopsies can Determine Latent Prostate Cancer Prevalence

To evaluate the usefulness of forensic autopsies in determining latent prostate cancer (PC) prevalence, we examined latent PC prevalence from autopsies and compared our findings between decedents with and without cancer. Data from forensic autopsies performed in Japan from 2004 to 2014 were obtained. For each prostate, histopathological examinations were performed in both the base and the apex sections. Three hundred and seventeen Japanese decedents were selected for analysis. The mean age of decedents was 56.4 ± 17.8 years (range, 14–94 years). Among this population, 39.4% died suddenly of disease and 60.6% died of external causes. Latent PC was identified in 45 (14.2%) decedents, who ranged from 27 to 93 years old (mean, 71.1 ± 12.9 years). The prevalence of clinically significant PC with a Gleason score of 7 or more was 8.8%, and the rate increased with age. Fifteen males had cancers other than PC. The prevalence of overall latent PC was significantly higher for those with cancer compared with those without (40.0% vs. 12.9%; P = 0.003). In this study, the use of forensic autopsy materials provided the opportunity to obtain a more accurate natural history of PC, as the decedents in this situation would have been more likely to have died suddenly while behaving as normal prior to death, and less likely to have been impacted by long-term medical interventions.     

Expedited, chemically enhanced sperm cell recovery from cotton swabs for rape kit analysis

This report focuses on the development of a method for chemically induced enhancement of cell elution and recovery from cotton swabs. The method exploits the exclusive use of detergents for intact cell removal, and can be utilized in conjunction with, or to circumvent, conventional differential extraction (DE). Samples treated with Sarkosyl (54.4 +/- 1.8%) and sodium dodecyl sulfate (SDS) (78.5 +/- 0.7%) yielded higher sperm cell recoveries than a conventional DE buffer (39.4 +/- 2.1%). The results indicated that the choice of detergent affected sperm cell yield, with anionic detergents having the greatest effect. Storage time of samples affected the concentration of detergent required for optimal sperm cell recovery, longer times requiring increased detergent concentrations. In addition, the extent of sperm cell lysis by proteinase K digestion was evaluated. The results indicate that the exclusive use of SDS enhances the release of sperm and epithelial cells from a cotton swab as compared with DE buffer, providing for a more effective DNA analysis.