Modulators of growth in primary culture of rat proximal tubular cells II

Summary: The present studies assessed the effects of manipulating extracellular sodium (Na) concentration and Na transport on cellular hypertrophy and hyperplasia in primary culture of rat proximal tubular cells. A concentration-dependent effect on thymidine incorporation and protein content was observed with cell culture media Na concentration of 130, 140 and 150 mmol/L. This effect was independent of osmolality (matched with mannitol) and no stimulatory effect occurred if choline was substituted for Na. Cells derived from sham-operated (Sx) animals exposed to a higher media concentration of Na (150 vs 140 mmol/L) had both stimulated thymidine incorporation to 186.8 ± 35.41% (P<0.05) and enhanced cell protein content to 134.7 ± 135% (P<0.05). This effect was more pronounced in cell cultures derived from unilaterally nephrectomized (Nx) animals, being 212.8 ± 31.5% (P<0.01) for thymidine incorporation (P<0.05 vs cells from sham-operated animals grown in high Na media) and 114.4 ± 3.2% (P<0.001) for protein content (P=0.11 vs sham-operated cells grown in similar conditions). the addition of 10^?4 mmol/L ethylisopropyl amiloride hydrochloride (EIPA) to Nx cells in a normal or high Na concentration media resulted in a decrease in cellular protein content to 82.6 ± 6.8% (P<0.05) and 85.5 ± 0.2% (P<0.0001) compared to respective controls. 10^?4 mol/L EIPA in media supplemented with insulin-like growth factor (IGF-1) blocked the proliferative response normally seen in response to this growth factor from 156.6 ± 13.7 to 27.5 ± 3.1% (P<0.0001) compared to control. However, the presence of EIPA did not abrogate the hypertrophic response elicited by IGF-1 (cell protein content 128.1 ± 13.1% of control with IGF-1 vs 124.9 ± 12.5 with IGF-1 and EIPA; P= n.s.). Addition of 10^?4 mol/L EIPA to 10% serum derived from either Sx or Nx animals blocked the growth response to the sera, limiting the cellular protein content to 76.6 ± 5.5% (P<0.0001) and 89.7 ± 4.4% (P<0.0001) and thymidine incorporation to quiescent levels of 0.2 ± 0.1% (P<0.0001) and 0.4 ± 0.1% (P<0.0001) compared to respective controls. In summary, rat renal proximal tubular cell growth is influenced by Na concentrations in the cell culture environment and inhibited in the presence of EIPA. This supports a role for altered epithelial transport in the cellular growth response to a number of stimuli.     

Reciprocity between tissue calmodulin and cAMP levels: modulation by excess zinc

Signal transduction of many intracellular events is initiated by a minute influx of calcium ions into the cells, resulting in the formation of calcium-calmodulin complex and cAMP. Because zinc appears to have an inhibitory effect on a number of tissue reactions, it is postulated that this occurs through modulation of intracellular calcium influx. To test the hypothesis that the inhibitory effects of zinc are mediated through the calcium calmodulin-cAMP pathway, zinc was administered by various routes to five groups of nude mice (control, intragastric, intraperitoneal. intradermal and oral groups), and calmodulin and cAMP concentrations were measured in the cytosol of epidermal cells. Calmodulin levels decreased significantly in the groups given intraperitoneal zinc (P>0.025) and intradermal zinc (P>0.001) injections. Significant elevations of cAMP levels were noted with intradermal zinc (P>0.025). Overall, the relationship between calmodulin and cAMP appeared to be inversely logarithmic, with the lowest calmodulin levels associated with the highest cAMP concentrations. In addition, there was a significant trend towards a smaller calmodulin/cAMP ratio in all zinc-treated groups, except the mice fed dietary zinc. These results appear to correlate with tissue zinc levels obtained with these various forms of zinc administration. Our results therefore indicate that there is a reciprocity between epidermal calmodulin and cAMP levels, which may be modulated by external factors such as zinc.     

真核表达人呼吸道合胞病毒F蛋白的纯化方法

目的 真核表达人呼吸道合胞病毒(human respiratory syncytial virus,SV)融合蛋白(fusion protein,),并完成蛋白纯化及纯度测定.方法 根据编码F蛋白的基因序列设计引物,CR方法扩增出3'端带His标签的F基因序列,克隆入pGEM-T-easy载体,经核酸序列分析后,进一步克隆到pcDNA3.1( )真核表达载体,限制性内切酶鉴定,用脂质体Lipofectamine2000转染COS-7细胞,2 h后再用Westem blot检测目的蛋白的表达.Ni柱亲和层析纯化COS-7细胞表达的F蛋白,高效毛细管电泳分析纯化后蛋白纯度.结果 核酸序列分析证实获得带His标签的RSV F基因序列,没有发生无义突变.转染COS-7细胞后,利用Western blot方法检测到F蛋白的特异性条带,纯度达99%以上.结论 初步建立了真核表达RSV F蛋白的纯化方法,为进一步优化RSV F蛋白制备条件及单克隆抗体及诊断试剂等研究奠定了基础.     

2017—2019年攀枝花市某医院心血管疾病住院患者疾病谱及住院费用分析

目的 分析心血管疾病住院患者疾病谱及住院费用,了解疾病构成的规律特点及经济负担,为心内科疾病诊治工作提供参考依据。方法 回顾性分析攀枝花市某三级医院2017—2019年心血管疾病住院患者资料,从医院相关管理信息系统中导出诊断为心血管疾病的病例条目、详细病历及病例住院费用等数据,采用描述流行病学分析方法对心血管疾病住院患者资料进行分析。结果 共纳入心血管疾病住院患者11 376例,2017—2019年分别有3 492、3 815、4 069例,分别占30.7%、33.5%、35.8%,病例数逐年上升。3年间心血管疾病住院患者的性别分布差异无统计学意义（P>0.05）,年龄分布差异有统计学意义（P<0.01）,心血管疾病住院患者发病呈年轻化趋势。病种均以冠心病和高血压所占比例最大,合计分别占当年心血管疾病住院患者的57.6%、55.5%,2019年则上升至62.8%。3年间心血管疾病住院患者有伴发疾病比例（27.6%、26.1%、25.4%）差异无统计学意义（P>0.05）。心血管疾病住院患者住院天数分布差异有统计学意义（P<0.01）,2019年的患者住院时间<7 d的比例有所上升。2017、2018、2019年该医院心血管疾病住院患者诊治自费费用分别为（5 925.6±285.7）、（5 904.2±264.3）、（5 890.9±230.8）元/人,差异有统计学意义（P<0.01）。结论 2017—2019年攀枝花市某医院心血管疾病住院患者病例数增加,住院患者发病年龄有年轻化趋势,主要病种仍以冠心病和高血压为主,住院天数及自费费用呈下降趋势。     

关于中药新药用饮片取样法的探讨

从药材及饮片（植物药）的包件取样、包件内取样及试验样品制备等方面对《中国药典》（Chinese Pharmacopoeia,ChP）、《欧洲药典》（European Pharmacopoeia,EP）、《英国药典》（British Pharmacopoeia,BP）及《美国药典》（United States Pharmacopoeia,USP）进行对比,结合新药用饮片检验的要求,就中药新药用饮片取样法存在的问题进行探讨,并提出以下建议：（1）相关部门组织专家起草中药新药用饮片取样的指导原则或技术要求。（2）暂时参照EP、BP的取样法对中药新药用饮片进行研究,并关注饮片取样对检验结果的影响。（3）根据试验样品的特点研究确定其处理方法。如饮片的不同药用部位或位置的质量差异大,可采用分层抽样的方法,减少取样误差。（4）加强药材的源头管理,减少药材质量差异。（5）在新药用饮片的生产过程中采用混合步骤提高同批饮片的同质性。（6）在ChP“中药材及饮片取样办法”中增加：待检批次饮片应具有较好的同质性;有些样品或试验需要采用更严格的取样方法,包括更多取样包件数或包件内取样量;此外,可对包件内抽取的位...     

人工心脏圆盘泵的流动特性研究

人工心脏（血泵）一直存在泵体对血细胞剪切力过大和流速过快容易引起溶血的问题。为了研究人体正常血压情况下,血泵内部剪切力和速度场的分布情况,选择圆盘泵叶轮代替传统离心泵叶轮,对两种模型进行数值计算,分析不同叶轮内部剪切力和速度场的分布规律。研究表明传统离心泵内部流速高,叶片表面剪切力大,对血细胞的伤害大。圆盘泵相比传统离心泵,剪切力更小,流场速度分布均匀,流速更小。和传统离心泵相比,不同转速下圆盘泵能降低溶血的发生率。圆盘泵叶片数为6片时,抗溶血性能更好。研究结果为血泵的优化提供理论依据。     

二氯乙酸对小鼠肝细胞DNA损伤效应研究

目的：研究二氯乙酸的DNA损伤作用。方法：小鼠肝细胞的体外、体内试验。体外试验，取小鼠肝细胞染毒，染毒’1h后彗星试验；体内试验，小鼠连续灌胃6d，每天一次，在第1、3、6d时各染毒3h后取其肝细胞进行彗星试验。荧光显微镜下观察、计数拖尾细胞的尾长。结果：无论是体外试验，还是体内试验，二氯乙酸均可导致肝细胞DNA损伤；体内试验结果显示．随染毒次数的增加，DNA损伤加重。结论：二氯乙酸是肝细胞的DNA损伤剂，致癌作用可能与其DNA损伤作用有关，属于遗传致癌物。     

肺癌细胞系内皮蛋白C受体的表达及其意义

目的 从细胞、蛋白和基因三个水平分析内皮蛋白C受体（endothelial protein C receptor，EPCR）在肺癌细胞系的表达。方法 以EA．hy926内皮细胞作阳性对照，人胚肺W138细胞作阴性对照，通过流式细胞术、West-ern．blot及半定量逆转录-聚合酶链反应分析EPCR在6种肺癌细胞系的表达水平。结果 6株肺癌细胞系均有EPCR的表达。高转移肺癌95D细胞表达最高。结论 EPCR的表达是肺癌细胞的重要特征，可能与肺癌增殖及转移有关。     

产科护理标识专科化管理的应用及效果

目的 探讨产科护理标识专科化管理的应用及效果.方法 通过规范产科病区各类基本标识的含义及作用,结合专科特点设计制作各类独立标识、组合标识、药物标识、仪器使用标识、人性化标识等用于临床护理工作.结果 实施产科护理标识专科化管理后,无1例因身份、管道识别发生的差错,无1例患者发生跌倒.实施前后分别对100例静脉滴注盐酸利托君注射液患者的液体滴速准确性进行比较,差异具有统计学意义(P＜0.05)；患者满意度由实施前的91.5％提高到98.0％.结论 产科护理标识的专科化管理能够有效警示护理工作重点,是人性化管理的有形展示,体现了护士的专业价值,加强了护理质量的全程控制,从而确保了护理安全.     

不同PCR联合用于鼠疫耶尔森菌的检测

目的 建立一种用4对不同引物快速鉴定鼠疫耶尔森菌的PCR方法。方法 分别采用4对针对V抗原，cafl，pla和inv4种基因的引物，在4只管中进行相同循环参数的扩增。结果 用这4对引物能特异性地扩增出鼠疫耶尔森菌217，478，524，295bp的目的DNA片段。结论 该法可用于鼠疫耶尔森菌简便、快速和准确的检测。