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.     

CDK4-pRB-E2F1通路是Aβ1-40诱导皮层神经元凋亡的可能途径

目的:探讨β淀粉样肽_1-40(beta-amyloidpeptide_1-40, Aβ_1-40)诱导大鼠皮层神经元凋亡的可能分子机制。方法:以40mg/L的Aβ_1-40诱导离体培养的大鼠皮层神经元凋亡, 流式细胞仪及免疫印迹方法检测细胞周期相关的周期依赖性激酶4(CDK4)、磷酸化的视网膜神经胶质瘤蛋白(pRB)水平;RT-PCR技术检测E2F1mRNA表达水平;荧光分光光度计检测半胱氨酸基天冬氨酸特异性蛋白酶3(caspase-3)活力;观察在Aβ_1-40诱导凋亡过程中是否伴随上述指标的变化。结果:①CDK4、磷酸化pRB水平在Aβ_1-40作用后2-4h显著升高;②Aβ_1-40孵育3h后皮层神经元E2F1基因表达上调;③Aβ_1-40作用12-24h后caspase-3活力明显升高。结论:CDK4-pRB-E2F1信号转导通路可能在Aβ_1-40诱导的大鼠皮层神经元凋亡中起主要作用。     

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...     

术中经胆囊管胆道造影的应用价值

目的探讨胆囊切除术时经胆囊管作胆道造影对胆道结石病人的应用价值。方法回顾分析442例患有胆囊胆道结石的病人作经胆囊管胆道造影的情况。结果 442例病人中,假阳性3例,假阴性1例,诊断准确率达99.1％。结论对胆囊胆道结石病人,特别是具有胆总管相对探查指征的病人,常规作术中经胆囊管胆道造影可降低胆道残余结石发生率,减少胆道损伤,避免不必要的胆总管探查。     

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。     

白首乌茎叶总黄酮对小鼠急性化学性肝损伤的保护作用

目的:研究白首乌茎叶总黄酮(TFC)对小鼠急性化学性肝损伤的保护作用。方法:腹腔注射四氯化碳(CCl4)制备小鼠急性肝损伤模型,检测血清丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)活性、肝脏重量指数;同时测定肝组织中丙二醛(MDA)、一氧化氮(NO)含量和超氧化物歧化酶(SOD)活性。结果:连续7d灌胃给予10,20mg/(kg·d)白首乌茎叶总黄酮,能明显抑制肝损伤小鼠血清ALT和AST活性的升高(P<0.01),减轻增加的肝脏重量指数,显著抑制肝组织中MDA和NO含量的升高及SOD活性的下降(P<0.01)。结论:白首乌茎叶总黄酮对CCl4引起的小鼠急性肝损伤具有保护作用,这种作用与其抗脂质过氧化有关。     

环氧合酶-2与肝细胞癌的关系

环氧合酶-2(cyclooxygenase-2)是前列腺素合成的限速酶，在许多恶性肿瘤中均有表达，并参与肿瘤的形成和复发转移。本文就COX-2的结构、在肝细胞癌中的表达及其抑制剂对肝细胞癌的作用作一综述。