穹窿主体蛋白对血管平滑肌细胞增殖的影响

目的:探讨穹窿主体蛋白(major vault protein,MVP)对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的影响?方法:分离培养野生型小鼠VSMCs,用血小板源性生长因子(PDGF-BB)或15% 胎牛血清(FBS)刺激VSMCs增殖,Western blot检测VSMCs的MVP表达水平,同时检测特异性分化标记物α-SMA和SM22α表达水平?然后用PDGF-BB刺激人主动脉平滑肌细胞(HAoSMCs),检测MVP的表达差异?再分别分离培养野生型小鼠和MVP敲除型小鼠的VSMCs,通过CCK8细胞增殖实验检测两者的增殖能力;同时用siRNA干扰的方法,使HAoSMCs中MVP低表达,检测其增殖能力的变化?结果:用PDGF-BB或15% FBS刺激VSMCs后,MVP表达水平呈浓度和时间依赖性下降,α-SMA和SM22α表达水平也相应降低?MVP缺失或低表达情况下,VSMCs的增殖能力均下降?结论:MVP在VSMCs增殖中起了重要作用?     

浙江省非小细胞肺癌患者EGFR基因与EML4-ALK融合基因突变的检测及其临床特征

目的:检测中国人非小细胞肺癌表皮生长因子受体(epidermal growth factor receptor,EGFR)基因和棘皮动物微管相关蛋白样-4与渐变性淋巴瘤激酶(EML4-ALK)融合基因的突变情况,同时分析这两种基因与非小细胞肺癌(non-small cell lung cancer,NSCLC)临床病理特征的关系?方法:采用PCR扩增和基因测序法检测252例NSCLCs EGFR基因外显子19和21的突变情况,real-time PCR法检测EML4-ALK融合基因的突变情况,分析突变与临床特征的关系?结果:252例非小细胞肺癌组织标本EGFR的突变率为38.8%(98例),其中19外显子突变率为15.8%(40例),21外显子突变率为23.0%(58例)?EGFR突变的患者多为女性?无吸烟史和腺癌(P < 0.05),与患者年龄无关(P > 0.05)?252例非小细胞肺癌组织标本中,EML4-ALK融合基因突变率4.7%(12例)?EML4-ALK基因突变患者多为女性和年龄较小者(P < 0.05),与患者吸烟史和病理类型无关(P > 0.05)?没有检测到EGFR和EML4-ALK的突变共存情况?结论:中国人中NSCLCs的EGFR突变率较高,在使用EGFR酪氨酸激酶抑制剂治疗前进行EGFR基因突变检测很有必要?EML4-ALK基因突变代表了NSCLC另一种分子亚型,这为临床NSCLC患者的治疗提供了一种新的选择方案?EML4-ALK融合基因突变与EGFR突变共存是罕见的现象?     

稳定表达小鼠PNPLA7的Huh7细胞系的构建

目的:构建稳定表达小鼠PNPLA7-Flag蛋白的人类肝癌细胞Huh7细胞系,观察小鼠PNPLA7在Huh7细胞中的过表达以及对Huh7细胞脂质含量的影响?方法:构建pCMV6 -PNPLA7-Flag真核表达载体,应用脂质体转染技术将该质粒导入Huh7细胞,随后通过G418筛选稳定表达的细胞系,运用Western blot方法检测PNPLA7蛋白表达;并通过油酸处理及油红染色观察过表达PNPLA7蛋白对Huh7细胞中脂质含量的影响?结果:成功获得2株稳定过表达PNPLA7-Flag的Huh7细胞系;Huh7细胞中过表达PNPLA7蛋白,在油酸处理后细胞内脂滴含量减少?结论:稳定表达小鼠PNPLA7的Huh7细胞系构建成功;小鼠PNPLA7在Huh7细胞中的表达会引起细胞中脂质含量减少,从而为后续研究PNPLA7蛋白在脂代谢通路的功能及机制提供可靠的细胞模型?     

人源抗H7N9血凝素中和抗体IgG的制备及鉴定

目的:将从全人源Fab噬菌体抗体库筛选获得的抗H7N9血凝素Fab抗体基因进行基因工程改造,以表达全分子抗H7N9血凝素中和抗体IgG,并验证其对H7N9型禽流感病毒的中和活性?方法:用H7N9型禽流感病毒血凝素筛选全人源Fab噬菌体抗体库,构建全分子IgG抗体重?轻链表达载体,共转染293 FreeStyle(293F)细胞,表达并纯化IgG抗体?应用ELISA及Western blot实验检测抗体免疫学活性,微量中和实验及鸡胚预防保护实验检测其中和活性,血凝抑制试验判断其结合血凝素亚基?结果:成功筛选出1株全人源抗H7N9血凝素Fab抗体基因并构建IgG真核表达载体?获得的全分子IgG抗体重?轻链序列正确,并可特异性结合H7N9血凝素?微量中和实验证明其对H7N9型禽流感病毒有良好中和活性,中和滴度为15.6 μg/mL?鸡胚预防保护实验显示抗体用量为200 μg时可达100%保护率?血凝抑制试验表明其结合血凝素HA2亚基?结论:制备的重组全人源全分子抗H7N9血凝素IgG抗体可特异性识别H7N9型禽流感病毒血凝素,对H7N9型禽流病毒有显著的中和作用,为进一步研发用于禽流感防治的抗体药物奠定基础?     

人源全分子抗c-Met 抗体的制备及对鼻咽癌细胞生物学特性的影响

目的:利用基因工程技术制备全分子人源抗 c-Met抗体,分析该抗体免疫学特性,并观察该全分子抗体对鼻咽癌细胞增殖?迁移和侵袭的影响?方法:设计引物扩增抗c-Met Fab抗体的重轻链可变区序列,将其分别克隆到真核表达载体pFUSE-CHIg-hG1 和 pFUSE-CLIg-hκ 中?转染真核细胞,表达产物用Protein A柱纯化?通过ELISA?Western blot和免疫荧光等方法鉴定该抗体的免疫学特性?CCK-8检测抗体对人鼻咽癌细胞增殖的抑制作用,划痕实验?Transwell侵袭实验分析抗体对人鼻咽癌细胞迁移和侵袭的影响?结果:成功重组全分子人源抗c-Met 抗体的表达载体,获得的全分子抗体保留了与抗原的特异性结合活性?CCK-8实验中结果显示全分子抗体能抑制c-Met阳性鼻咽癌细胞CNE2和HONE1的增殖?细胞划痕和Transwell侵袭实验结果显示全分子抗体能抑制c-Met阳性细胞CNE2和HONE1的迁移和侵袭能力?结论:本研究成功制备人源全分子抗c-Met抗体,该抗体有明显的中和活性,为探索鼻咽癌分子靶向治疗奠定了基础?     

医学院校图书馆创客空间的构建

从创客空间的概念和发展现状出发,分析了我国医学院校图书馆构建创客空间的意义,探讨了医学院校图书馆构建创客空间的主要形式,包括构建新型学习空间,知识共享空间和协作交流空间。并提出了构建我国医学院校图书馆创客空间的建议。     

Association ofmicroRNA polymorphisms withthe risk ofhead andneck squamous cell carcinoma ina Chinese population:a case-control study

Background:MicroRNA (miRNA) polymorphisms may alter miRNA-related processes, and they likely contribute to cancer susceptibility. Various studies have investigated the associations between genetic variants in several key miR-NAs and the risk of human cancers; however, few studies have focused on head and neck squamous cell carcinoma (HNSCC) risk. This study aimed to evaluate the associations between several key miRNA polymorphisms and HNSCC risk in a Chinese population. Methods:In this study, we genotyped ifve common single-nucleotide polymorphisms (SNPs) in several key miRNAs (miR-149 rs2292832,miR-146a rs2910164,miR-605 rs2043556,miR-608 rs4919510, andmiR-196a2 rs11614913) and evaluated the associations between these SNPs and HNSCC risk according to cancer site with a case–control study including 576 cases and 1552 controls, which were matched by age and sex in a Chinese population. Results:The results revealed thatmiR-605 rs2043556 [dominant model: adjusted odds ratio (OR) 0.71, 95% conif-dence interval (CI) 0.58–0.88; additive model: adjusted OR 0.74, 95% CI 0.62–0.89] andmiR-196a2 rs11614913 (domi-nant model: adjusted OR 1.36, 95% CI 1.08–1.72; additive model: adjusted OR 1.28, 95% CI 1.10–1.48) were signiifcantly associated with the risk of oral squamous cell carcinoma (OSCC). Furthermore, when these two loci were evaluated together based on the number of putative risk alleles (rs2043556 A and rs11614913 G), a signiifcant locus-dosage effect was noted on the risk of OSCC (Ptrend<0.001). However, no signiifcant association was detected between the other three SNPs (miR-149 rs2292832,miR-146a rs2910164, andmiR-608 rs4919510) and HNSCC risk. Conclusion:Our study provided the evidence thatmiR-605 rs2043556 andmiR-196a2 rs11614913 may have an impact on genetic susceptibility to OSCC in Chinese population.     

Bisphenol A disrupts glucose transport and neurophysiological role of IR/IRS/AKT/GSK3β axis in the brain of male mice

Bisphenol A (BPA), one of the most prevalent chemicals for daily use, was recently reported to disturb the homeostasis of energy metabolism and insulin signaling pathways, which might contribute to the increasing prevalence rate of mild cognitive impairment (MCI). However, the underlying mechanisms are remained poorly understood. Here we studied the effects of low dose BPA on glucose transport and the IR/IRS/AKT/GSK3β axis in adult male mice to delineate the association between insulin signaling disruption and neurotoxicity mediated by BPA. Mice were treated with subcutaneous injection of 100 μg/kg/d BPA or vehicle for 30 days, then the insulin signaling and glucose transporters in the hippocampus and prefrontal cortex were detected by western blot. Our results showed that mice treated with BPA displayed significant decrease of insulin sensitivity, and in glucose transporter 1, 3 (GLUT1, 3) protein levels in mouse brain. Meanwhile, hyperactivation of IR/IRS/AKT/GSK3β axis was detected in the brain of BPA treated mice. Noteworthily, significant increases of phosphorylated tau and β-APP were observed in BPA treated mice. These results strongly suggest that BPA exposure significantly disrupts brain insulin signaling and might be considered as a potential risk factor for neurodegenerative diseases.     

靶向成纤维细胞活化蛋白放射性探针研究进展

成纤维细胞活化蛋白（FAP）是癌症相关成纤维细胞和活化成纤维细胞的重要生物标志物,在许多肿瘤和纤维化组织的活化成纤维细胞上高表达,而在正常组织和非恶性肿瘤中基本不表达。因此,FAP已经成为肿瘤等疾病的诊断和治疗的优良靶标。基于FAP抑制剂（FAPI）的PET显像和内放射疗法已成为包括癌症、纤维化等多种疾病在内的新型诊疗手段,以介绍FAP介导疾病发生发展的机制及其作为靶标的诊疗意义为开端,对¹²⁵I、⁶⁸Ga、⁶⁴Cu等多种放射性核素标记靶向FAP探针的结构演进、显像、体内生物分布及其药代动力学性质进行了系统总结,归纳了改善探针的药代动力学性质和靶标亲和力的相关策略,包括采用方酰胺连接子、使用白蛋白结合剂加以修饰、开发双靶向探针,对未来靶向FAP新型放射性探针的开发和经典探针的临床应用提出了一些参考建议。     

炎症性肠病症状量表评估炎症性肠病疾病活动度的临床价值研究

背景 国外研究显示炎症性肠病症状量表（IBDSI）可用于炎症性肠病（IBD）患者进行疾病活动的自我评价,但该量表目前尚未在国内得到验证。 目的 探究IBDSI评估IBD疾病活动度的价值。 方法 选取2020年4月至2021年8月在南京医科大学附属淮安第一医院住院治疗的IBD患者102例,其中溃疡性结肠炎（UC）患者59例,克罗恩病（CD）患者43例。采用改良的Mayo评分评估UC病活动度,克罗恩疾病活动指数（CDAI）评分评估CD疾病活动度。患者入院后次日清晨填写由本院消化内科医生翻译的中文版IBDSI。IBDSI及各子量表评分与改良的Mayo评分,CDIA评分及实验室检查指标的相关性分析采用Spearman秩相关分析;绘制IBDSI及其各子量表评分（包括肠道症状、腹部不适、疲劳、肠道并发症、全身并发症）评估UC、CD疾病活动度的受试者工作特征曲线（ROC曲线）。 结果 59例UC患者改良的Mayo评分：临床缓解1例,轻度8例,中度46例,重度4例;IBDSI评分（26.27±15.16）分。43例CD患者CDAI评分：缓解期11例,轻度8例,中度22例,重度2例;IBDSI评分（26.95±16.19）分。Spearman秩相关分析结果显示,UC患者IBDSI总分、肠道症状评分、腹部不适评分、疲劳评分与改良的Mayo评分呈正相关（P<0.05）。CD患者IBDSI总分、肠道症状评分、腹部不适评分、疲劳评分、全身并发症评分与CDAI评分呈正相关（P<0.05）。IBDSI及肠道症状、腹部不适、疲劳子量表评估UC疾病活动度的ROC曲线下面积（AUC）分别为0.829、0.836、0.710、0.786（P<0.05）;IBDSI及肠道症状、腹部不适、疲劳、全身并发症子量表评价CD疾病活动度的AUC分别为0.911、0.806、0.785、0.867、0.734（P<0.05）。 结论 IBDSI及各子量表评分与UC和CD的疾病活动度存在一定相关性,有助于初步判断患者疾病严重程度。