临床医学专业“5+3”一体化本科全程导师制创新人才培养的实践与探索

以培养创新型人才为目标，大连医科大学制定实施了“5+3”创新人才培养改革方案，以导师制培养为载体，在医学本科教育全过程中，制定分阶段创新能力培养体系，涵盖课程、讲座、实验设计、论文等基本科研能力训练，强化本科生科研能力培养。通过对首届“5+3”学生阶段性培养成果的统计学分析发现，“5+3”学生发表中文期刊、SCI，主持国家级创新项目、省级创新项目的比例均显著高于普通5年制学生，差异具有统计学意义（P<0.05）。虽然实施过程中存在一些问题和不足，但以导师制为核心的科研基础训练对提高学生科研思维和创新能力效果显著，对培养医学创新型人才具有可实施性。     

Inhibitors of the Hepatitis C Virus Polymerase; Mode of Action and Resistance

The hepatitis C virus (HCV) is a pandemic human pathogen posing a substantial health and economic burden in both developing and developed countries. Controlling the spread of HCV through behavioural prevention strategies has met with limited success and vaccine development remains slow. The development of antiviral therapeutic agents has also been challenging, primarily due to the lack of efficient cell culture and animal models for all HCV genotypes, as well as the large genetic diversity between HCV strains. On the other hand, the use of interferon-α-based treatments in combination with the guanosine analogue, ribavirin, achieved limited success, and widespread use of these therapies has been hampered by prevalent side effects. For more than a decade, the HCV RNA-dependent RNA polymerase (RdRp) has been targeted for antiviral development. Direct acting antivirals (DAA) have been identified which bind to one of at least six RdRp inhibitor-binding sites, and are now becoming a mainstay of highly effective and well tolerated antiviral treatment for HCV infection. Here we review the different classes of RdRp inhibitors and their mode of action against HCV. Furthermore, the mechanism of antiviral resistance to each class is described, including naturally occurring resistance-associated variants (RAVs) in different viral strains and genotypes. Finally, we review the impact of these RAVs on treatment outcomes with the newly developed regimens.     

miR-21对视网膜母细胞瘤细胞增殖的影响及作用机制

目的　探讨MicroRNA-21（miR-21）对视网膜母细胞瘤(RB)细胞增殖的影响及作用机制。方法　采用Real-time PCR技术检测miR-21在正常视网膜组织和确诊RB组织中的表达情况；然后在转染的基础上运用MTT检查RB细胞存活率、流式细胞仪检测细胞凋亡率。Western blot法检测与细胞凋亡相关蛋白PDCD4、Bax、Bcl-2的表达。结果　与正常视网膜组织miR-21表达 （0.703±0.071）相比，RB组织miR-21为高表达（2.214±0.162），差异有统计学意义（P<0.01）。在Weri-Rb-1细胞中，与NC组(2.245±0.213)相比，miR-21抑制剂转染后明显降低了miR-21的表达水平，miR-21 inhibitor组为0.683±0.075,差异有统计学意义 （P<0.01）。两组细胞转染后24 h、48 h、72 h、96 h，MTT测定法检测细胞活力结果显示:两组24 h的A值比较，差异无统计学意义 （P>0.05），miR-21 inhibitor 组在 48 h、72 h、 96 h的A值均低于 NC 组，差异均有统计学意义 （均为P<0.01）。流式细胞术检测结果显示:NC组凋亡细胞在总细胞中百分比为（3.045±0.301）%和（4.832±0.493）%，miR-21 inhibitor组凋亡细胞在总细胞中百分比为（2.593±0.257）%和（40.167±4.014）%，miR-21 inhibitor组Weri-Rb-1细胞的凋亡率明显高于miR-21 NC组(P<0.01)。Western blot检测结果显示:NC组PDCD4表达（0.192±0.045）相比miR-21 inhibitor组（0.683±0.091）表达明显减少，NC组Bax的蛋白表达水平（0.143±0.036）相比miR-21 inhibitor组（1.192±0.054）也明显减少，差异均有统计学意义（P<0.01)，NC组Bcl-2蛋白表达（0.864±0.038）相比miR-21 inhibitor组（0.257±0.026）明显增多，差异有统计学意义（P<0.05)。结论　miR-21是RB的促癌基因，miR-21抑制剂可以通过降低miR-21表达抑制肿瘤细胞的增殖、促进细胞凋亡，这一过程与PDCD4、Bax、Bcl-2等凋亡相关蛋白有关。     

lncRNA RP11-86H7.1在川崎病诊断中的临床意义

目的：探讨lncRNA RP11-86H7.1在川崎病（KD）患者血清中的表达及其与临床病理特征及预后的关系。方法：筛选KD特异相关的循环lncRNA，分KD治疗前患儿组、KD治疗后患儿组、普通发热患儿组及健康儿童组，采用qPCR检测各组血清lncRNA RP11-86H7.1相对表达。分析血清lncRNA RP11-86H7.1相对表达与KD临床病理特征间关系；绘制ROC曲线，分析血清lncRNA RP11-86H7.1表达水平对KD的诊断效能。结果：KD急性患儿组血清lncRNA RP11-86H7.1相对表达量高于各对照组（P＜0.05）；年龄和性别比例与低表达组比较差异无统计学意义（P＞0.05）；qPCR发现lncRNA RP11-86H7.1在KD急性期患儿血清中表达水平明显高于KD恢复期、健康儿童及发热儿童组，差异均有统计学意义（P＜0.05）。结论：血清lncRNA RP11-86H7.1在KD患者中表达上调，其可作为KD早期诊断和评估预后的潜在的生物标志物。     

miR-200、 miR -155及血管新生因子与复发性流产的相关性分析

目的 研究miR-200、miR-155及血管新生因子与原因不明复发性流产(unexplained recurrent spontaneousabortion,URSA)的相关性分析。 方法 选择2015年3月—2018年1月在青岛市妇女儿童医院妇产科就诊的URSA患者作为URSA组、要求终止妊娠的正常早孕患者作为对照组,检测绒毛组织中微小RNA(microRNA, miR)miR-200、miR-155、血管内皮生长因子(vascular endothelial growth factor,VEGF)、可溶性FMS样酪氨酸激酶1(soluble FMS like tyrosine kinase-1,sFlt-1)的表达量及血清中VEGF、sFlt-1的含量,对miR-200、miR-155靶向结合VEGF、sFlt-1进行生物信息学分析。 结果 URSA组的绒毛组织中miR-200(1.78±0.32 vs. 0.91±0.15)、sFlt-1(1.87±0.35 vs. 1.06±0.21)的相对表达量及血清中sFlt-1的含量[(12.39±2.31)ng/ml vs. (6.51±0.95)ng/ml]均高于对照组,差异有统计学意义(均P<0.05)。绒毛组织中miR-155相对表达量(0.60±0.10 vs. 0.93±0.16)、VEGF mRNA相对表达量(0.59±0.09 vs. 1.02±0.16)及蛋白表达量(0.62±0.07 vs. 1.04±0.18)、血清中VEGF的含量[(601.25±94.39)ng/ml vs. (935.12±132.47)ng/ml]低于对照组,差异有统计学意义(均P<0.05);URSA组患者绒毛组织中miR-200的表达量与血清中VEGF的含量、绒毛组织中VEGF的表达量均呈负相关,绒毛组织中miR-155的表达量与血清中sFlt-1的含量和绒毛组织中sFlt-1的表达量均呈负相关;miR-200、miR-155分别靶向结合VEGF、sFlt-1基因的3’UTR。 结论 miR-200表达增多、miR-155表达减少与URSA发生有关,miR-200靶向VEGF、miR-155靶向sFlt-1是介导该过程的可能机制。     

Design and synthesis of novel organometallic complexes using boronated phenylalanine derivatives as potential anticancer agents

Drug design and discovery studies are important because of the prevalence of diseases without available medical cures. New anticancer agents are particularly urgent because of the high mortality rate associated with cancer. A series of mononuclear gold (III) and platinum (II) complexes based on boronated phenylalanine (BPA) were designed and synthesized using 4,4’-dimethyl-2,2’-dipyridyl (L1) or 1,10-phenanthroline-5,6-dion (L2) ligands to obtain promising anticancer drug candidates. Proton nuclear magnetic resonance, infrared, mass spectrometry, and elemental analyses were utilized for chemical characterizations. Cell viability, cancer cell colony formation, endothelial tube formation, and cytoskeleton staining assays were performed using A549 lung adenocarcinoma and human umbilical vein endothelial cells (HUVECs) to investigate preliminary pharmacological activities. L1-based platinum (II) complex (BPA-L1-Pt) was the most promising complex, and has similar activity with the approved chemotherapy drug cis-platinum. Half maximal inhibitory concentration values for BPA-L1-Pt were 9.15 µM on A549s and 16.61 µM on HUVECs; the values for cis-platinum were 5.24 µM on A549s and 23.14 µM on HUVECs. Consequently, further synthesis studies should be performed to boost the cancer cell selectivity feature of BPA by varying metal and ligand types.