Sertad1 Induces Neurological Injury after Ischemic Stroke via the CDK4/p-Rb Pathway

SERTA domain-containing protein 1 (Sertad1) is upregulated in the models of DNA damage and Alzheimer’s disease, contributing to neuronal death. However, the role and mechanism of Sertad1 in ischemic/hypoxic neurological injury remain unclear. In the present study, our results showed that the expression of Sertad1 was upregulated in a mouse middle cerebral artery occlusion and reperfusion model and in HT22 cells after oxygen-glucose deprivation/reoxygenation (OGD/R). Sertad1 knockdown significantly ameliorated ischemia-induced brain infarct volume, neurological deficits and neuronal apoptosis. In addition, it significantly ameliorated the OGD/R-induced inhibition of cell viability and apoptotic cell death in HT22 cells. Sertad1 knockdown significantly inhibited the ischemic/hypoxic-induced expression of p-Rb, B-Myb, and Bim in vivo and in vitro. However, Sertad1 overexpression significantly exacerbated the OGD/R-induced inhibition of cell viability and apoptotic cell death and p-Rb, B-Myb, and Bim expression in HT22 cells. In further studies, we demonstrated that Sertad1 directly binds to CDK4 and the CDK4 inhibitor {"type":"entrez-nucleotide","attrs":{"text":"ON123300","term_id":"2214659616"}}ON123300 restores the effects of Sertad1 overexpression on OGD/R-induced apoptotic cell death and p-Rb, B-Myb, and Bim expression in HT22 cells. These results suggested that Sertad1 contributed to ischemic/hypoxic neurological injury by activating the CDK4/p-Rb pathway.     

七星瓢虫基础研究现状

七星瓢虫对蚜虫等农林业中的重要害虫有较强的控制作用,利用七星瓢虫进行控害的生物防治技术正在逐步推广.本文就七星瓢虫的生物学特性、捕食、对农药的反应以及人工饲养等方面进行综述:七星瓢虫幼虫随龄期增长逐渐出现黄斑,温湿度对生长发育影响较大,以成虫越冬,有滞育现象.七星瓢虫捕食性强,捕食对象较多,包括多种蚜虫、木虱、蚧虫、粉...     

Rab31 promotes the invasion and metastasis of cervical cancer cells by inhibiting MAPK6 degradation

Persistent infection with high-risk human papillomavirus (HPV) is the main risk factor for cervical cancer. Our mass spectrometry data showed that the Ras-associated binding protein Rab31 was upregulated by HPV; however, little is known regarding the role of Rab31 in the metastasis of cervical cancer cells. In this study, we showed that Rab31 was highly expressed in cervical cancer tissues and cells, and both HPV E6 and E7 promoted the expression of Rab31. Rab31 knockdown inhibited while Rab31 overexpression promoted the migration and invasion capabilities of cervical cancer cells. Additionally, Rab31 knockdown inhibited the epithelial-mesenchymal transition (EMT) and cytoskeletal rearrangement in cervical cancer cells. Furthermore, Rab31 interacted with mitogen-activated protein kinase 6 (MAPK6), and Rab31 knockdown inhibited the expression of MAPK6, which was mainly localized in the cytoplasm. More importantly, Rab31 knockdown promoted and Rab31 overexpression inhibited MAPK6 degradation. Accordingly, MAPK6 overexpression restored the decreased migration potential caused by Rab31 knockdown. Finally, a xenograft mouse model showed that Rab31 knockdown in cervical cancer cells led to reduced tumor growth and impaired lung and liver metastasis in vivo. In conclusion, Rab31 plays a crucial role in cervical cancer metastasis by inhibiting MAPK6 degradation. Thus, Rab31 may serve as a novel therapeutic target to manage cervical cancer.     

(4-Piperidinylphenyl)aminoethyl amides as a novel class of non-covalent cathepsin K inhibitors

A series of (4-piperidinylphenyl)aminoethyl amides based on dipeptide anilines were synthesized and tested against cathepsin K, cathepsin L and cathepsin B. These new non-covalent inhibitors exhibited single-digit nM inhibition of the cysteine proteases. Compounds 3 and 7 demonstrated potency in both mouse and human osteoclast resorption assays.     

A human TERT C-terminal polypeptide sensitizes HeLa cells to H2O2-induced senescence without affecting telomerase enzymatic activity

We have constructed a 27-kDa hTERT C-terminal polypeptide (hTERTC27) devoid of domains required for telomerase activity and demonstrated that it is capable of nuclear translocation/telomere-end targeting. Here we showed that expression of a low level of hTERTC27 renders hTERT positive HeLa cells sensitive to H(2)O(2)-induced oxidative stress and subsequent cell senescence. The senescence-associated gene, the cyclin/cdk inhibitor p21(Waf1), was up-regulated. This occurs without changing the expression of endogenous hTERT, causing significant telomere shortening or inhibiting telomerase activity. Results from this study suggest for the first time that in addition to telomerase activity, the C-terminus of hTERT also plays a role in hTERT-mediated cellular resistance to oxidative stress.     

High-level expression of a truncated wall-associated protein A from the dental cariogenic Streptococcus mutans

Streptococcus mutans plays a primary role in the formation of dental caries. Previously, in our laboratory, an S. mutans genomic library was prepared, and the wapA gene was cloned into the shuttle vector, pSA4/4B2. To generate overexpression of wapA and to facilitate efficient purification of the WapA protein for use as an immunogen, an expression vector with the strong tac promoter was used. In order to answer questions regarding the optimization of solubility and expression based on gene size or the hydrophobicity of the protein product, 12 truncated constructs of the wapA gene were prepared using PCR. The truncated products were subcloned into the pGEX-6P-1 glutathione S-transferase (GST) fusion vector and expressed in E. coli BL21. The fusion proteins were analyzed by SDS-PAGE and confirmed by analysis with anti-GST and anti-WapA antibodies. Our study suggests that abrogation of the wapA promoter is necessary for expression of this gene in this expression system. Deletion of the signal peptide and the hydrophobic C terminus of WapA increased expression compared with the full-length construct, and truncation at the protease cleavage site of the C-terminal region greatly increased the stability of the protein without a loss in reactivity with the anti-WapA antibody. Western immunoblot analysis with anti-WapA antiserum clearly showed that the majority of the epitopes of the GST-WapA fusions are located in the N-terminal region of WapA. The immunogenicity of the various WapA fusion products is being examined in mice and rats to further map the immunologically dominant regions of the protein. This method effectively increased the expression of WapA and should contribute to the further understanding of gene expression of E. coli, as well as aid in the characterization of this protein for future immunologic evaluation.     

柑桔原生质体辐射诱变筛选抗寒再生植株

用１１６１Ｃ／ｋｇ软Ｘ射线辐射“Ｐａｇｅ”桔柚（ＣｉｔｒｕｓｒｅｔｉｃｕｌａｔａＢｌａｎｃｏ×Ｃ．ｇｒａｎｄｉｓＯｓｂ．ｃｖ．Ｐａｇｅ）胚性愈伤组织分离的原生质体,经－１１℃低温选择后,将存活的原生质体培养再生植株。研究表明,“Ｐａｇｅ”桔柚胚性愈伤组织原生质体在４１２８Ｃ／ｋｇ辐射剂量处理后几乎不能恢复分裂;２０６４Ｃ／ｋｇ辐射时分裂频率低,即使分裂也难以发育为胚状体;而１１６１Ｃ／ｋｇ辐射后,经－１１℃低温处理仍有１０２％～１６９％的分裂频率,且部分细胞团能发育为胚状体。辐射的原生质体培养成胚状体后,经低温筛选,在改良的生芽培养基和生根培养基中萌发成为植株。再生植株中检测出２株的叶片原生质体的低温半致死温度（ＬＴ５０）分别比对照低１．９６℃和１６８℃。     

糖类在植物组织培养中的效应

糖类是影响植物组织培养成功与否的关键之一。迄今为止,已用于植物组织培养的糖类有５０多种。在植物体细胞组织培养中,蔗糖一直作为标准碳源,然而其他糖类物质如：葡萄糖、麦芽糖和山梨醇对植物体细胞培养也产生了一定的影响。在植物花药培养中,蔗糖较好,但应注意麦芽糖对花药培养的促进作用。