84例抗凝药物致不良反应的帕累托图分析

目的： 分析抗凝药物不良反应（ADR）发生的规律、特点,探讨ADR发生与变化的可能原因及影响因素,为临床应用提供参考。方法： 对上海交通大学附属第一人民医院2012年1月-2018年12月上报至国家药品不良反应监测系统的84例抗凝药物所致ADR进行帕累托图分析。结果： 84例抗凝药物ADR报告中,男性比例较高为46例（54.76%）;60岁以上人群ADR发生例次较多,有62例（73.81%）;新的ADR报告4例（4.76%）,严重的ADR报告19例（22.62%）;发生ADR的抗凝药物共4个种类,其中低分子肝素类药物最多（41.84%）,其次为维生素K拮抗剂类（27.55%）;以血液系统损害最为常见（74.23%）;以药师上报为主。结论： 抗凝药物引发ADR与多种因素有关,临床用药时应全方位监控,以减少ADR的发生,确保用药安全。     

神经上皮细胞转化基因1对细胞辐射敏感性的影响及其机制探讨

目的 探讨神经上皮细胞转化基因1(Net1)对细胞辐射敏感性的影响及相关的分子作用机制.方法 运用实时荧光定量PCR检测辐射后细胞中Net1基因表达水平的变化;采用RNAi干扰技术抑制细胞中Net1的表达,用克隆形成率分析细胞的辐射敏感性;利用免疫共沉淀技术发现Net1的结合蛋白.结果 电离辐射损伤后,细胞中的Net1 mRNA水平显著上升(t=-10.52,P＜0.05);与对照组相比,siRNA沉默细胞中的Net1表达后明显增加了细胞的辐射敏感性(t=15.31、11.65,P＜0.05);无论在正常状态下还是在细胞受到辐照后,Net1都能与非同源末端连接修复蛋白Ku70、Ku80和DNA-PKcs结合.结论 Net1对细胞的辐射防护作用可能是通过与非同源末端连接修复蛋白相互作用来调控辐射损伤修复实现的.     

老年人股骨颈骨折股骨头置换术中短柄假体的有限元分析

目的针对老年人股骨颈骨折后假体置换术,使用有限元分析的方法对股骨短柄假体与常规股骨假体的生物力学特性进行对比。方法利用薄层CT资料建立股骨三维有限元模型,利用我院股骨中空多孔股骨假体相关参数建立两种髋关节股骨假体三维有限元模型;应用Ansys 5.7有限元分析软件考察两种假体植入后的应力分布,并进行比较。结果短柄假体所受最大mises应力较常规股骨假体明显减小;力学传导模式仍为远端应力集中,最大mises应力仍出现在张力侧。结论老年人股骨颈骨折关节置换术中短柄假体力学特性优越。     

Ezrin shRNA联合HSP70对骨肉瘤细胞凋亡和增殖的影响

 目的 探讨Ezrin基因沉默联合热休克蛋白（heat shock protein, HSP）70诱导的免疫杀伤对骨肉瘤细胞增殖和凋亡的影响。方法 采用人成骨肉瘤MG63细胞系作为研究对象,将人HSP70和Ezrin-shRNA的DNA片段分别克隆至含CMV和pU6双启动子的表达载体pGFP-V-RS中构建含HSP70和Ezrin-shRNA的真核表达载体pGFP-V-RS-shRNA和pGFP-V-RS-shRNA-HSP70。重组质粒分别转染入细胞,筛选出稳定表达的细胞克隆。荧光显微镜观察细胞形态及转染效果;荧光定量RT-PCR及蛋白印迹western blot检测稳定转染细胞株Ezrin和HSP70基因及蛋白水平的变化;采用MTT、流式细胞术检测细胞增殖、凋亡能力变化,Western blot检测凋亡及周期相关蛋白表达量变化;MTT法检测HSP70刺激产生特异性细胞毒性T淋巴细胞（CTL）对靶肿瘤细胞的杀伤作用。结果 荧光图像及基因和蛋白表达分析证实,首次成功构建同时沉默Ezrin和过表达HSP70的特异性载体。较单纯沉默Ezrin,同时过表达HSP70会在一定程度上减弱沉默Ezrin蛋白促进细胞凋亡和抑制增殖的效果,但与对照细胞相比,M63细胞凋亡率仍明显上升,自11.01%±0.22%上升至24.28%±0.50%,增殖速度则自395.14%±2.24%减少至310.00%±2.83%。另外,Western blot检测发现Ezrin-shRNA可促进凋亡基因Bax的表达,相反可降低抗凋亡基因Bcl-2和细胞周期蛋白Cyclin D1的表达。而Ezrin-shRNA/HSP70组较Ezrin-shRNA组促Bax表达和降低Bcl-2、细胞周期蛋白Cyclin D1表达有所减弱,但较阴性对照组仍有明显效果。MG63细胞的CTL细胞毒杀伤效应显示各效靶浓度下CT+IL-2+HSP70组的杀伤活性高于CT+IL-2组,最高达56.33%±1.95%。结论 同时沉默Ezrin、过表达HSP70既可以促进骨肉瘤细胞凋亡抑制其增殖,并利用HSP70诱导的CTL,增强对靶肿瘤细胞的杀伤作用。     

儿童股骨远端骨肉瘤保留骨骺的定制肿瘤型假体重建术

 目的 介绍儿童股骨远端骨肉瘤保留骨骺的定制肿瘤型假体重建术,并探讨其工作原理、手术操作技巧、早期临床疗效以及并发症的防治。方法 2012年8月至2013年7月期间,应用肿瘤瘤段骨切除、保留骨骺的定制肿瘤型假体重建术治疗股骨远端骨肉瘤的儿童患者3例,均为男性,年龄8岁、9岁和15岁。术前给予新辅助化疗1~2周期,化疗结束后根据X线、CT 和MR等检查结果评价化疗疗效,对于化疗效果好且符合保留骨骺手术条件者采用此术式治疗。首先通过CT、MRI确定病变范围,根据影像学检查结果利用计算机辅助定制保留骨骺的肿瘤型假体以及模具,待假体定制完毕后行肿瘤瘤段骨切除、保留骨骺的定制肿瘤型假体重建术,术后指导患者进行功能锻炼,切口愈合1周后给予术后的规范化化疗,并进行长期随访。 结果 3例患者手术时间分别为3 h、4 h和6 h,术中出血量分别为300 ml、500 ml和2 200 ml。对3例患者术后随访时间 为12~24个月,根据美国骨肿瘤学会评分系统（Musculoskeletal Tumor Society,MSTS）,术后3个月功能评分分别为24分、26分和13分,短期随访显示患者肢体功能良好。1例患者出现假体感染,经保守治疗（抗炎补液、切口换药等）无效后行大腿截肢术,余2例患者未出现假体松动等其他并发症。2例患者双下肢长度相差均< 2 cm。结论 通过严格掌握保留骨骺保肢手术的适应证,配合术前及术后的新辅助化疗,保留骨骺的定制肿瘤型假体重建术为儿童股骨远端骨肉瘤的保肢治疗提供了新的选择方案,其疗效安全、可靠,且具有手术操作简单、手术时间短、术后恢复快等优点,但长期疗效尚需进一步观察。     

Characteristics of spermatogonial stem cells derived from neonatal porcine testis

The aim of this study was to isolate and characterise porcine spermatogonial stem cells (PSSCs). The putative porcine germline stem cells from testis were isolated successfully by an improving way of enrichment with lymphocyte separation medium (LSM). Results from RT‐PCR analyses showed that PSSCs were positive for OCT4, SOX2, NANOG, PGP9.5, c‐MYC, KEL4 and PRDM‐14 which are multipotent stem cell markers. At the protein level, the results of immunofluorescence analyses showed that PSSCs were positive for OCT4, PGP9.5, SOX2 and CD29. We successfully differentiated these PSSCs into adipocytes and muscle cells and then defined their characteristics, including morphology, surface stem cell markers, and mechanical properties. But the experiment of teratoma formation was negative. The results indicated the PSSCs could be multipotent. Atomic force microscopy was used to characterise the morphological and mechanical properties of undifferentiated PSSCs, as well as the differentiated adipocytes and muscle cells, which could be potentially useful for distinguishing PSSCs from differentiated cells.     

Growth and Aging of Proximal Femoral Bone: A Study With Women Spanning Three Generations

Osteoporotic hip fracture is a serious clinical event associated with high morbidity and mortality. Understanding femoral growth patterns is important for promoting bone health in the young and preventing fractures in later life. In this study, growth patterns of areal bone mineral density (aBMD) and geometric properties of the proximal femur were measured by dual‐energy X‐ray absorptiometry. They were studied in 251 girls from premenarche (11.2 ± 0.7 years) to late adolescence (18.3 ± 1.1 years) and compared with their premenopausal mothers (n = 128, aged 44.9 ± 4.1 years) and postmenopausal grandmothers (n = 128, aged 70.0 ± 6.3 years). Hip axis length (HAL) was the first to reach peak growth velocity (?10.5 months before menarche), followed by neck diameter (ND) and neck cross‐sectional area (CSA), (?7.1 and ?4.1 months before menarche, respectively). Both neck‐shaft angle (NSA) and aBMD of neck and total hip peaked at menarche. At 18 years (7‐year follow‐up), girls already had higher femoral neck aBMD but similar HAL and NSA compared with their mothers. Grandmothers had the longest HAL, narrowest NSA, widest ND but lowest aBMD and CSA. Hip strength index (HSI), an index of femoral neck strength during a fall, dropped rapidly after menarche in girls but thereafter remained relatively constant. Grandmothers had lower HSI than either mothers or girls. In conclusion, differences in proximal femoral bone mass and structure in adulthood are largely established before menarche, indicating that heritable factors are responsible for most of the individual variance. The development of geometric properties precedes aBMD in puberty, resulting in relatively constant hip strength after menarche. This asynchronous growth leads to adaptation of bone strength to the imposed loads, avoiding fractures in a biologically efficient manner. Both deterioration of aBMD and inadequate compensatory change in bone geometry after menopause contribute to the increased fracture risk later in life. © 2014 American Society for Bone and Mineral Research.