Chemical Composition of Chinese Pyrola Herb Volatile and the Effects on the Osteoblast Proliferation

目的分析鹿衔草挥发油的化学成分,研究其对体外培养成骨细胞增殖的影响。方法采用水蒸气蒸馏法提取鹿衔草挥发油,GC-MS检测化学成分;鹿衔草挥发油干预成骨细胞系ROS17/2.8后,采用MTT法检测成骨细胞增殖,流式细胞术检测成骨细胞增殖周期,实时荧光定量PCR检测成骨细胞PCNA mRNA表达。结果一定浓度的鹿衔草挥发油能促进成体外培养成骨细胞增殖,处于增殖周期的成骨细胞比例明显增加,且成骨细胞的增殖细胞核抗原(Proliferating Cell Nuclear Antigen,PCNA)mRNA表达显著提高。结论鹿衔草挥发油通过上调PCNA表达,加速成骨细胞增殖周期进程,从而促进成骨细胞增殖。Objective To analyze the chemical composition of Chinese pyrola herb volatile and study its effects on the in vitro cultured osteoblast proliferation. Methods The water vapor distillation was used to extract Chinese pyrola herb volatile and GC- MS was to determine the chemical composition. After intervention of Chinese pyrola herb volatile on ROS17 /2. 8,MTT method was adopted to determine the osteoblast proliferation,the flow cytometry was to determine the osteoblast proliferating cycle and RT- PCR was to determine the expression of PCNA mRNA. Results Chinese pyrola herb volatile of a certain of concentration promoted the proliferation of in vitro cultured osteoblast cells. The percentage of osteoblast cells in the proliferation cycle was increased apparently and the expression of PCNA mRNA was improved significantly. Conclusion Chinese pyrola herb volatile accelerates the progression of osteoblast proliferating cycle through up- regulating PCNA expression so as to accelerate osteoblast proliferation.国家自然科学基金(81473706);; 福州市卫生系统科技计划项目(2013-S-wq10);; 福建省中医药科研项目(wzgs201307

Study of the effect of Strong-bone granules on the differentiation of ROS1728 osteoblasts with silenced ER expression

目的探讨健骨颗粒对成骨细胞中ERalpha介导的TERT信号通路的调控作用。方法采用雌激素受体拮抗剂ICI182780(Faslodex)阻断成; 骨细胞中雌激素受体alpha(ERalpha)的表达,建立ER抑制的大鼠成骨细胞株ROS1728细胞模型,采用酶联免疫吸附法(ELISA)检测成; 骨细胞液中碱性磷酸酶(alkaline phosphatase,ALP) 、骨钙素(osteocalcin,BGP); 、Ⅰ型胶原(collagen I,Col Ⅰ)的含量。采用实时荧光定量SYBR; GREEN法检测ERE、ERalpha、c-MYCmRNA的表达。采用Western Blot检测TERT、ERalpha、c-; MYC蛋白的表达。结果ELISA法检测结果显示:随着干预时间的延长,培养液中的ALP、BGP、ColⅠ的含量逐渐上升。其中对照组3种信号因子的含; 量最高血,雌激素组次之,健骨颗粒组再次之,模型组最低,各组比较差异有统计学意义(P < 0.05),; 4组TERT、ERalpha、c-MYCmRNA及蛋白表达量情况以对照组的蛋白表达含量最高,雌激素组次之,健骨颗粒组再次之,模型组最低。各组比较; 均有统计学意义(P < 0.05); 。结论雌激素介导TERT信号通路及其相关因子与成骨细胞分化的关系密切,而补肾健脾中药健骨颗粒可通过雌激素介导TERT信号通路促进成骨细胞分化。Objective To investigate the effect of Strong-bone granules on the; regulation of TERT signaling pathway mediated by ER alpha in; osteoblasts. Methods The estrogen receptor antagonist ICI182780; (Faslodex) was used to inhibit the expression of ER in osteoblasts and; to establish the ER-silenced model of rat osteoblast cell line ROS1728.; Serum ALP,BGP,and Col I were determined using enzyme-linked; immunosorbent assay. The mRNA expression of ERE,ERalpha,and c-MYC was; determined using real time quantitative SYBR GREEN assay. The protein; expression of ERE,ERalpha,and c-MYC was detected using Western blotting.; Results The results of ELISA showed that the content of ALP,BGP,and Col; in the culture medium increased gradually with the prolonging of; intervention time. The levels were the highest in the control group,then; followed in estrogen group,Strong-bone granules group,and the model; group,and the difference among the groups was significant (P < 0.05).; The mRNA and protein expression of TERT,ER alpha,and c-MYC was the; highest in the control group,then followed in estrogen group,Strong-bone; granules group,and the model group,and the difference among the groups; was significant (P < 0.05). Conclusion The estrogen mediated TERT; signaling pathway and its related factors are closely related to the; differentiation of osteoblasts. Strong-bone granules promote osteoblast; differentiation through estrogen mediated TERT signaling pathway.国家自然科学基金; 福建省卫生厅中医课题; 福州市卫生系统科技项

Effects of Jiangu Granules on the proliferation of osteoblasts through G_1/S phase cell-cycle regulated proteins

目的观察健骨颗粒对成骨细胞G_1/S期调控的影响,探讨健骨颗粒促进成骨细胞增殖的作用机制。方法制备健骨颗粒血清组、模型血清组和雌二醇血清组。采用酶消化法培养SD大鼠成骨细胞,健骨颗粒含药血清干预,以模型血清和雌二醇血清为对照。运用流式细胞术检测成骨细胞增殖周期,荧光定量PCR法检测成骨细胞G_1/S期调控蛋白Cyclin E、CDK2、p21和转录因子E2F-1 m RNA的表达。结果 15%的雌二醇血清与健骨颗粒血清干预48 h,成骨细胞增殖速度均明显快于模型血清组(P<0.01);G_0/G_1期成骨细胞比例明显降低(P<0.01),S期、G_2/M期细胞比例及增殖指数则明显高于模型血清组(P<0.01);与模型血清组比较,雌二醇血清组与健骨颗粒血清组能提高成骨细胞Cyclin E、CDK2及转录因子E2F-1 m RNA的表达(P<0.01),而降低p21的表达(P<0.01)。结论健骨颗粒通过调节成骨细胞G_1/S期调控机制,推进成骨细胞顺利通过G_1/S检测点,促进成骨细胞增殖。Objective To observe the effects of Jiangu Granules on the G_1/S phase cell-cycle regulated proteins of osteoblasts, so as to reveal the mechanism of Jiangu Granules in promoting osteoblasts proliferation. Methods The Jiangu Granules-serum group, model-serum group and estradiol-serum group were prepared. Osteoblasts of SD rats were cultivated by enzymatic digestion and intervened with Jiangu Granules-serum, model-serum and estradiol-serum(as control) respectively. The cell cycles were analyzed by flow cytometry, while the fluorescence quantitative PCR was applied to measure cyclin E, CDK2, p21 and E2F-1 m RNA. Results The cell proliferation rates of osteoblasts intervened with estradiol serum and Jiangu Granules-serum for 48 hours were faster than that with the same concentration model-serum(P < 0.01). Compared with the model-serum group, the proportion of osteoblasts in the G_0/G_1 phase were significantly reduced after intervention with 15% of estradiol serum and Jiangu Granules-serum for 48 hours(P < 0.01),while the proportion of cells in S phase, G_2/M phase and proliferation index was much higher(P < 0.01). The expression of cycling E, CDK 2 and transcription factor E2F-1 m RNA of osteoblasts in the Jiangu Granules-serum group and the estradiol-serum group was higher than that of model-serum group(P < 0.01), while the expression of p21 m RNA was lower than the model-serum group(P < 0.01). Conclusion Jiangu Granules-serum can adjust the G_1/S phase cellcycle regulated mechanism in osteoblast, so as to push the cells passing G_1/S checkpoint and accelerate the proliferation of osteoblast.国家自然科学基金资助项目(81473706

Thermokinetics Study on Antibacterial Activity of Schiff Base Co(Ⅲ)Complex

用微量热法测定了大肠杆菌及其在Co(Ⅲ ) -SG配合物作用下的热谱曲线 ,计算了速率常数k ,传代时间G、抑制I、总产热量Q、单个细菌的产热量Q0 和每分钟单个细菌的产热量 Q0 ,建立了部分代谢参数之间的关系 ,探讨了大肠杆菌在不同温度和Co(Ⅲ ) -SG作用下的代谢抑制 ,发现可用t、tr、tg和 Q0 定量表征在不同温度下大肠杆菌的生长代谢和药物的抗菌活性 .　The power -time curves of E .coli reacting with Co(Ⅲ)-SG at different temperature has been determined by microcalorimtric method The multiplication rate constant k , generation time G, bacterial growth inhibition ratio I , total thermogenetic quantity Q , the heat quantity of a single bacterium Q0 and the heat quantity of a single bacterium per minute Q have also been calculated according to the power -time curves .On the basis of k ～ T data , the formula lnk ～ 1/ T has graphically obtained and activation energy Ea of bacterial growth and pre -exponetial factor A'have been calculated.by graphing the parameters of E .coli growth metabolism , the relationships of tr ～ k , tg ～ k and t ～ k have been derived .The paper provides a discussion about the growth metabolism of E .coli reacted upon by Schiff base Co (Ⅲ)complex at different temperatures, and it is found that tr , tg , t and Q0 can be used to characterize bacterial growth metabolism and the antibacterial activity of drugs at different tmperature

非晶硅薄膜太阳电池的数值优化研究

基于Sentaurus TCAD数值分析平台,采用非晶硅的DOS模型对禁带中缺陷态进行表征,建立a-Si：H薄膜太阳电池的二维数值模型。对P-I-N结构的非晶硅太阳电池的本征区、P型区、N区以及P/I界面的特性进行研究,得到参数与薄膜太阳电池性能之间的关系。通过电池物理和结构参数的优化,在界面处引入ZnO作为反射层,优化得到太阳电池填充因子为74.7%,AM1.5下光电转换效率为10.1%,表明采用TCAD数值仿真可有效用于非晶硅太阳电池本征参数和反射层的优化设计,提高电池转换效率

利用分析Unigene在转录组中表达模式的方法拼接盐角草铵转运基因/Assembling of an ammonium transporter gene in Salicornia europaea by expression pattern analysis of Unigene in transcriptome[J]

RNA-seq技术能够全面快速地获得物种在某一状态下的转录本序列信息,但测序并组装后的大量Unigene往往不包含完整ORF (Open reading frame).转录组库具有一定的冗余性,存在着属于同一个转录本的Unigene,这些Unigene因为无重叠区不能拼接而存在转录组库中.基于这种情况,为了拼接铵转运蛋白家族Unigene,首先挑选注释为AMT (Ammonium transporter)且ORF不完整的所有Unigene(5条),通过分析Unigene在4个转录组的表达模式,其中2条Unigene (Uni4和Uni5)具有相同的表达模式,推测可能来自同一转录本.然后通过NCBI blastx将这2条Unigene与参考物种的AMT蛋白质比对,确定其在转录本的位置及序列相互间没有交叠(如果两条编码序列相互交叠则不能组成同一个转录本).结果发现Uni4和Uni5分别位于参考转录本5 '端和3'端位置,因此假定它们属于同一个转录本,中间空缺约120 bp未知序列.通过试验验证,分别在Uni4和Uni5上设计单正向引物和单反向引物,PCR扩增得到约800 bp片段,将其测序并与两条Unigene比对,证实Uni4和Uni5属于同一转录本且获得了缺失的未知序列.最终拼接得到1 667 bp序列,包含l 482 bp完整ORF,编码494个氨基酸,通过系统进化分析将其归类为amt1亚家族,命名为Seamt1.生物信息学手段预测SeAMT1蛋白与已知的其他物种AMT性质相似.本研究采用转录组Unigene表达模式聚类的方法挖掘潜在的同一转录本Unigene,并且通过另外两组Unigene检验了该方法的可行性.这一便捷方法有助于转录组中Unigene的延伸和拼接,有助于完整ORF的获得及后期基因功能研究

盐胁迫下盐角草 Na ＋／H ＋转运基因表达分析/Expression analysis of Salicornia europaea Na+/H+antiporters under salt stress[J]

Na＋/H＋逆向转运蛋白具有调节细胞内离子浓度及维持pH值稳定的作用，是植物抵御盐胁迫的重要因子。从盐角草RNA-Seq数据中筛选Na＋/H＋逆向转运蛋白序列，利用生物信息学手段，拼接得到5条Na＋/H＋逆向转运蛋白完整cDNA序列，通过与NCBI已有序列比对分析，将其命名为SeNHX1、SeNHX3、SeNHX4、SeNHX5和SeN-haD。考察盐角草Na＋/H＋逆向转运蛋白在两种盐分变化情况下的表达变化：1）从无盐处理转移到200 mM NaCl处理；2）从200 mM NaCl培养基转移到无盐培养基。结果表明：SeNHX4的表达量非常低，在地下部几乎检测不到；SeNhaD在地上部的表达量是地下部的2倍左右，说明NhaD主要在盐角草地上部发挥作用；SeNHX1、SeNHX3和SeNHX5的表达量明显高于其他两个基因，对盐角草的耐盐机制起到更大的作用，并且SeNHX1和SeNHX5受盐分诱导表达，表达量与盐浓度呈正相关，可能在盐角草耐盐调控网络中发挥重要作用。综上，NHX基因家族和NhaD的表达量受到基质中盐分的调控作用，其表达在盐分处理或者盐分去除1～3d后发生变化。研究结果有助于阐明盐角草NHX基因家族和NhaD对盐分的响应特点