我院护士在职教育的体会

本文针对不同层次护理人员制定不同的培训目标和计划,采取岗前培训,岗位培训(专科培训、在职分层培训、护理人才培训),自学等形式,开展护士在职教育,不断扩充更新知识,以适应现代护理工作要求。     

力学刺激促进骨髓基质干细胞体外软骨分化

目的探讨离心力刺激对猪骨髓基质干细胞(BMSCs)体外成软骨分化的作用,以及对三维支架上构建组织工程化软骨的影响,明确力学刺激与细胞分化及组织形成的关系,为体外软骨构建提供适当参数。方法抽取8周龄猪髂嵴骨髓,应用贴壁法分选单个核细胞,体外培养扩增后获得第2代BMSCs,以5.0×107/cm3的细胞密度接种到聚羟基乙酸(PGA)制成的圆柱形三维支架上,7d后分成4组,在不同的力学条件及诱导条件下培养。8周后取材,行相关检测。结果力学诱导组形成的组织呈白色,细腻光泽,形状规则,体积无明显改变,有良好的弹性和硬度,并具有典型的软骨陷窝结构和大量软骨特异性细胞外基质,Ⅱ型胶原及丰富的聚合蛋白多糖(GAG)成分。GAG含量为(6.0±1.2)mg/g,抗压强度为(2.2±0.8)kPa,弹性模量为(7.4±1.6)kPa。各项指标均明显优于其他各组。结论力学刺激有利于促进BMSCs成软骨分化,并在三维支架材料上构建组织工程化软骨。     

雌二醇对不同性质的乳腺原代细胞增殖影响的体外研究

目的了解在体外条件下不同浓度的雌二醇对不同性质的乳腺组织原代细胞增殖的影响。方法用胶原酶消化法获取乳腺癌患者的各类乳腺组织原代细胞，并用MTT法检测乳腺癌组织、癌旁组织和正常乳腺组织原代细胞在体外培养下，给予不同浓度雌二醇时细胞的生长增殖情况。结果雌激素在体外对雌激素受体（ER）表达阳性的乳腺癌组织及癌旁组织原代细胞有明显的刺激生长作用；对正常乳腺组织细胞的促生长作用相对较弱，而且与肿瘤组织的ER表达情况无关。结论当体内的雌二醇水平≥10μmol／L时，乳腺癌的发病风险和复发风险会显著增加。     

In vivo quantification of fluorescent molecular markers in real-time: A review to evaluate the performance of five existing methods

With the advent of molecular-targeted fluorescent markers, there is a renewed interest in fluorescence quantification methods that are based on continuous wave excitation and multi-spectral image acquisition. However, little is known about their in vivo quantification performance. We reviewed the performance of five selected methods by analytically describing these and varying input parameters of irradiance, excitation geometry, collection efficiency, autofluorescence, melanin content, blood volume, blood oxygenation and tissue scattering using optical properties representing those for human skin. We identified one method that corrects for variations in all parameters. This requires image acquisition before and after marker administration, under identical geometry. Hence, it is suited for applications where the site of interest can be relocated (e.g. anaesthetized animals and dermatology). For applications where relocation is not possible, we identified a second method where the uncertainty in the fluorescence signal was ±20%. Hence, use of these methods can substantially aid in vivo fluorescence quantification compared to use of the raw fluorescence signal, as this changed by more than 3 orders of magnitude. Since these methods can be computed in real-time, they are of particular interest for applications where direct feedback is critical, as diagnostic screening or image-guided surgery.     

加减下瘀血汤对低密度及氧化型低密度脂蛋白诱导小鼠肾足细胞血管内皮生长因子分泌的干预作用

目的：观察脂质低密度（LDL）及氧化型低密度脂蛋白（OX-LDL）对体外培养小鼠足细胞分泌血管内皮生长因子（VEGF）的影响及加减下瘀血汤的保护作用.方法：观察不同浓度下LDL和OX-LDL对小鼠肾足细胞分泌VEGF的影响,再用不同浓度加减下瘀血汤含药血清进行干预,分别用RT-PCR及ELISA检测足细胞分泌VEGF的变化.结果：LDL和OX-LDL可使肾足细胞分泌VEGF明显增高,且有浓度依赖性.中药加减下瘀血汤含药血清的干预可以明显抑制脂质诱导肾足细胞VEGF的高表达.结论：LDL和OX-LDL可以通过促进足细胞分泌VEGF增加而加重脂质肾毒性,而中药加减下瘀血汤可能通过抑制脂质诱导足细胞分泌过多VEGF,从而保护足细胞的脂质损伤.     

辛伐他汀对人低密度脂蛋白氧化修饰影响的实验研究

目的研究辛伐他汀在体外对低密度脂蛋白氧化修饰的影响.方法以低密度脂蛋白氧化修饰为模型,以硫代巴比妥酸反应物质(TBARS)生成量及相对电泳迁移率(REM)为指标,研究了辛伐他汀对铜离子(Cu2 )诱导低密度脂蛋白氧化修饰的抑制作用.结果随辛伐他汀浓度从1～10μmol/L的增加,TBARS值分别减少67.3%,73.9%,81.9%,REM值减少48.3%,55.2%,58.6%,呈浓度及时间依赖关系.其中10μmol/L辛伐他汀可几乎完全抑制低密度脂蛋白氧化.结论辛伐他汀在体外能明显抑制Cu2 诱导的低密度脂蛋白氧化修饰.     

17β-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响

目的研究17β-雌二醇(17β-E2)对子宫内膜异位症(内异症)患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响,探讨Wnt/β-catenin信号通路在介导雌激素促进内异症发生发展的作用。方法体外分离培养内异症患者在位子宫内膜间质细胞。用不同浓度17β-E2处理子宫内膜间质细胞48 h;此后选用10-10mol/L 17β-E2处理子宫内膜间质细胞12、24和48 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western blotting)检测17β-E2处理前后子宫内膜间质细胞β-catenin mRNA和蛋白的表达水平。同法分析雌激素受体拮抗剂ICI182,780(10-6mol/L)对17β-E2促进β-catenin mRNA和蛋白表达的影响。免疫组织化学染色观察17β-E2作用后β-catenin在子宫内膜间质细胞中的定位。结果17β-E2能明显促进内异症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白的表达,并呈剂量和时间依赖性,于10-10mol/L作用48 h最明显。雌激素受体拮抗剂ICI182,780能明显抑制17β-E2对子宫内膜间质细胞β-catenin mRNA和蛋白的表达。免疫组织化学染色发现17β-E2能促进β-catenin在子宫内膜间质细胞核内的表达。结论雌激素可能通过激活Wnt/β-catenin信号通路促进内异症在位子宫内膜的异位种植。     

四种中药对骨愈合过程中相关基因表达的影响

目的:探索中药水蛭、海螵蛸、阿胶、骨碎补在骨折愈合过程中的干预作用,了解它们各自的调节靶点,探索建构其基因组学的途径。方法:通过在大鼠胫骨打孔的方法建立单因素干扰模型,并将300只大鼠随机分为正常组、模型组和给药组(分别用4种中药给药),每组50只,分别在实验的第4、7、14、21、28天不同时间点采用原位杂交方法对各类mRNA的变化进行动态观察,分析骨愈合过程中Ⅰ、Ⅱ、Ⅲ型前胶原mRNA、转化生长因子TGF-β1mRNA、骨形态发生蛋白BMP-2mRNA以及血管内皮生长因子VEGF-mRNA的表达情况。结果:不同中药对不同基因的作用不同,作用的时间点不同,作用强度也存在差异。其中海螵蛸在骨折早期对Ⅰ、Ⅲ型前胶原mRNA、VEGF-mRNA、BMP-2mRNA的表达升高,后期Ⅱ、Ⅲ型前胶原mRNA表达水平下降,VEGF-mRNA、TGF-β1mRNA表达量维持于较高水平;骨碎补组较模型组在BMP-2mRNA、TGF-β1mRNA、Ⅰ型前胶原mRNA的表达上差异有显著性统计意义;阿胶对骨愈合早、中期Ⅰ、Ⅱ、Ⅲ型前胶原mRNA和TGF-β1mRNA的表达与模型组比较差异存在显著性统计意义;水蛭对VEGF-mRNA的表达具有一定的促进作用。结论:海螵蛸、水蛭对血管形成有促进作用,阿胶、骨碎补和海螵蛸对骨折软骨形成早期具有促进骨诱导的作用,并对成骨细胞的增殖及合成活性有较大影响。     

ReProComet: a new in vitro method to assess DNA damage in mammalian sperm.

The increasing request of chemical safety assessment demands for the validation of alternative methods to reduce the resort to animal experimentation. Methods that evaluate reproductive toxicity are among those requiring the largest use of animals. Presently, no validated in vitro alternative exists for the assessment of reproductive toxicity. Mammalian sperm are sensitive targets of DNA-reactive chemicals, which form premutagenic adducts. Here, we propose a new method based on comet assay to detect DNA damage induced by potential germ cell mutagens in bull sperm available from assisted reproduction practices. In somatic cells, chemical-induced adducts can be revealed by comet assay that detects DNA breaks produced during adduct repair. Mature sperm, however, are devoid of repair enzymes, and adducts are processed only after fertilization. For this reason, comet assay is not sensitive to detect DNA lesions induced in sperm by most chemicals. To overcome such limitation, we developed a modified comet assay based on the addition of a protein extract from HeLa cells to agarose-embedded sperm on microscopic slides. To test the method, sperm were treated in vitro with methyl methanesulfonate (MMS) or melphalan (MLP) and comet assay was conducted both with and without protein supplementation. No effect of MMS or MLP was detected without protein supplementation; on the contrary, a clear-cut dose-dependent effect was measured after addition of the cell extract. These results represent a proof of concept of a novel in vitro mutagenicity test on sperm that could offer a promising approach to complement previously validated in vivo germ cell genotoxicity assays.     

关于我国体外诊断试剂监督管理改革的建议

本文综述了美国、欧盟、日本、中国目前对体外诊断产品（试剂）监督管理的现状以及我国存在的问题。并根据我国具体情况提出了对体外诊断试剂监督管理的改革建议。