17个新的C2H2型锌指基因片段的分离与克隆

按照Ｃ２Ｈ２型锌指基因保守结构域的ＤＮＡ序列设计一对简并引物,以人基因组ＤＮＡ为模板进行ＰＣＲ同源扩增,将由此获得的锌指基因片段为探针,从人胎肾、骨骼肌、骨骼组织的ｃＤＮＡ分子库中筛选到２２个Ｃ２Ｈ２型锌指蛋白ｃＤＮＡ片段,经国际ＮＣＢＩ数据库查询检索,其中１７个为新的锌指基因片段。对从胎肾ｃＤＮＡ分子库中分离到的Ｋ３－４和Ｋ５－１２克隆进行了表达谱分析,发现Ｋ３－４在肾脏中的表达量明显高于其他几     

Effects of rare earth elements on telomerase activity and apoptosis of human peripheral blood mononuclear cells

To study the effects of rare earth exposure on human telomerase and apoptosis of mononuclear cells from human peripheral blood (PBMNCs). The blood contents of 15 rare earth elements, including La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, and Y, were measured by inductively coupled plasma-mass spectrometry. Telomeric repeat amplification protocol assay and flow cytometer analysis were carried out to analyze the telomerase activity and apoptosis of PBMNCs, respectively. The total content of rare earth elements in the blood showed significant differences between the exposed group and the control group. The rare earth exposure increased the telomerase activity and the percentages of cells in the S-phase and the G₂/M phase in PBMNCs, but it had no effect on the apoptotic rate of PBMNCs. Under the exposure to lower concentrations of rare earth elements, the telomerase activity of PBMNCs in the exposed group was higher than that of the control group, and there was no effect on the apoptotic rate of PBMNCs, but promoted the diploid DNA replication and increased the percentages of G₂/M- and S-phase cells.     

Expression of Oct4 in HCC and modulation to wnt/β-catenin and TGF-β signal pathways

Hepatocellular carcinoma (HCC) is considered as a disease of dysfunction of the stem cells. Studies on stem cells have demonstrated that Oct4 plays a pivotal role in embryo regulation. In order to understand the role of Oct4 in HCC and the relationship among Oct4 and wnt/β-catenin and TGF-β signal pathways, we have detected the expression of Oct4, Nanog, Sox2, STAT3 as well as the genes in wnt/β-catenin, and TGF-β families in HCC cell lines and in tumor specimens from HCC patients. The authors found that Oct4 was expressed in all of the four HCC cell lines and the tumor specimens from HCC patients. Some other genes were also expressed in them with different level including Nanog, Sox2, STAT3 and TCF3, wnt10b, β-catenin, ELF, Smad3 and Smad4. The ability of the clone formation and migration of the HepG2 decreased after Oct4 was knockdowned. Silencing of Oct4 and TCF3 in HCC cell line HepG2 revealed that there were complicated relationships among Oct4, wnt/β-catenin family and TGF-β family genes. Knockdowning Oct4 reduced the expression of TGF-β family genes ELF, Smad3, Smad4 and wnt/β-catenin family genes, wnt10b, and β-catenin but increased TCF3. In reverse, knockdowning TCF3 led to the increased expression of Oct4 and TGF-β family genes. In conclusion, the expression of Oct4 in HCC may play an important role as in stem cell. Because Oct4 improves not only the function of wnt/β-catenin, but also the TGF-β signal pathways, the significance of its expression in HCC might be more complicated than we evinced before.     

一种新型改良硫氧还蛋白融合表达体系的建立及cathelicidin家族Lf-CATH2的表达

通过改良硫氧还蛋白融合表达体系,原核表达cathelicidin家族抗菌肽Lf-CATH2。首先在Lf-CATH2基因上游加入凝血酶位点,并去除p ET32α载体的凝血酶序列和S标签序列,构建优化的Lf-CATH2-p ET32α-TS载体,于大肠杆菌中表达。产物融合蛋白经凝血酶切割释放Lf-CATH2,纯化后进行抗菌活性检测。结果表明改良的硫氧还蛋白融合表达体系显著提高酶切效率达37%,Lf-CATH2在新体系中获得了可溶性高表达,且保留了抗菌活性。因此该新型硫氧还蛋白融合表达体系,有望为cathelicidin家族及其他阳离子活性肽提供更好的原核表达载体工具。     

干旱,半干旱地区作物育种的困惑与出路

粮食问题主要取决于一年生谷类作物产量。作物产量低而不稳的原因主要是病虫害及各种胁迫生境,其中干旱缺水为最大的产量限制因素,提高作物生产力的途径有二：其一是改善作物的生长环境,其二是通过育种手段选育在各肿胁迫环境中具有优良表现的基因型（品种）。矮秆化育种手段使水肥充裕区小麦产量有显著的提高,是通过提高收获指数获得的。干旱、半干旱地区育种却未能获得显著效果,要提高干旱、半干旱地区小麦育种的成效,对干旱     

Exosomes derived from umbilical cord mesenchymal stem cells alleviate viral myocarditis through activating AMPK/mTOR‐mediated autophagy flux pathway

Human umbilical cord mesenchymal stem cell‐derived exosomes (hucMSC‐exosomes) have been implicated as a novel therapeutic approach for tissue injury repair and regeneration, but the effects of hucMSC‐exosomes on coxsackievirus B3 (CVB3)‐induced myocarditis remain unknown. The object of the present study is to investigate whether hucMSC‐exosomes have therapeutic effects on CVB3‐induced myocarditis (VMC). HucMSC‐exosomes were identified using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and Western blot. The purified hucMSC‐exosomes tagged with PKH26 were tail intravenously injected into VMC model mice in vivo and used to administrate CVB3‐infected human cardiomyocytes (HCMs) in vitro, respectively. The effects of hucMSC‐exosomes on myocardial pathology injury, proinflammatory cytokines and cardiac function were evaluated through haematoxylin and eosin (H&E) staining, quantitative polymerase chain reaction (qPCR) and Doppler echocardiography. The anti‐apoptosis role and potential mechanism of hucMSC‐exosomes were explored using TUNEL staining, flow cytometry, immunohistochemistry, Ad‐mRFP‐GFP‐LC3 transduction and Western blot. In vivo results showed that hucMSC‐exosomes (50 μg iv) significantly alleviated myocardium injury, shrank the production of proinflammatory cytokines and improved cardiac function. Moreover, in vitro data showed that hucMSC‐exosomes (50 μg/mL) inhibited the apoptosis of CVB3‐infected HCM through increasing pAMPK/AMPK ratio and up‐regulating autophagy proteins LC3II/I, BECLIN‐1 and anti‐apoptosis protein BCL‐2 as well as decreasing pmTOR/mTOR ratio, promoting the degradation of autophagy flux protein P62 and down‐regulating apoptosis protein BAX. In conclusion, hucMSC‐exosomes could alleviate CVB3‐induced myocarditis via activating AMPK/mTOR‐mediated autophagy flux pathway to attenuate cardiomyocyte apoptosis, which will be benefit for MSC‐exosome therapy of myocarditis in the future.