Stable kinetochore–microtubule interactions depend on the Ska complex and its new component Ska3/C13Orf3

Ska1 and Ska2 form a complex at the kinetochore–microtubule (KT–MT) interface and are required for timely progression from metaphase to anaphase. Here, we use mass spectrometry to search for additional components of the Ska complex. We identify C13Orf3 (now termed Ska3) as a novel member of this complex and map the interaction domains among the three known components. Ska3 displays similar characteristics as Ska1 and Ska2: it localizes to the spindle and KT throughout mitosis and its depletion markedly delays anaphase transition. Interestingly, a more complete removal of the Ska complex by concomitant depletion of Ska1 and Ska3 results in a chromosome congression failure followed by cell death. This severe phenotype reflects a destabilization of KT–MT interactions, as demonstrated by reduced cold stability of KT fibres. Yet, the depletion of the Ska complex only marginally impairs KT localization of the KMN network responsible for MT attachment. We propose that the Ska complex functionally complements the KMN, providing an additional layer of stability to KT–MT attachment and possibly signalling completion of attachment to the spindle checkpoint.     

一步3’RACE快速构建鸡MnSOD全长cDNA克隆Rapid

本研究尝试将触减 PCR与3’ cDNA末端快速扩增（rapid amplification of cDNA ends,RACE）技术进行结合,仅用一条特异性引物和一条通用引物,成功地实现了从3’末端cDNA库对鸡含锰超氧化物歧化酶（manganese-containing superoxide dismutase,MnSOD）全长cDNA的一步3’RACE快速构建。与常规使用的末端PCR或亚克隆方法相比,该法具有快速、省时、经济和特异性好的优点。Abstract: RACE(rapid amplification of cDNA ends) is a popular technique to rapidly obtain the full-length cDNA. After obtaining the 3’cDNA and 5’cDNA fragments with a overlapped region by 3’RACE and 5’RACE, the full-length cDNA could be generated by end-to-end PCR or subcloning. In this study, 3’RACE combined with touch-down PCR was successfully used for the rapid construction of full-length MnSOD cDNA of chickens. Compared with the conventional end-to-end PCR or subcloning, this method, called one-step 3’RACE, is fast, economical and highly specific. It especially fits the rapid construction of full-length cDNA by RACE method.     

The EPSPS gene flow from glyphosate-resistant Brassica napus to untransgene B. napus and wild relative species Orychophragmus violaceus

Gene flow from transgenic plants to compatible wild relatives is one of the major impediments to the development of the culture of genetically engineered crop plants. In this work, the flow of EPSPS (conferring resistance to glyphosate) gene of transgene Brassica napus toward the untransgene B. napus and wild relative species Orychophragmus violaceus in an open field (1 ha) was studied. The data related to only the 2004 and 2005 autumn season on one location of southwest of China. Pollen dispersal and fertilization of the target plants were favored and a detailed analysis of the hybrid offspring was performed. In field, the data studied show that the gene flow frequency was 0.16% between GM and non-GM B. napus at a distance of 1 m from the transgenic donor area. The crosspollination frequency was 0.05% between GM and non-GM B. napus at a distance of 5 m from the transgenic donor area. At a distance of 10 m, no crosspollination was observed. According to the results of this study, B. napus transgene flow was low. However, the wild relative species O. violaceus could not be fertilized by the transgenic pollen of B. napus, no matter what the distance was.     

Epitope Tags beside the N-Terminal Cytoplasmic Tail of Human BST-2 Alter Its Intracellular Trafficking and HIV-1 Restriction

BST-2 blocks the particle release of various enveloped viruses including HIV-1, and this antiviral activity is dependent on the topological arrangement of its four structural domains. Several functions of the cytoplasmic tail (CT) of BST-2 have been previously discussed, but the exact role of this domain remains to be clearly defined. In this study, we investigated the impact of truncation and commonly-used tags addition into the CT region of human BST-2 on its intracellular trafficking and signaling as well as its anti-HIV-1 function. The CT-truncated BST-2 exhibited potent inhibition on Vpu-defective HIV-1 and even wild-type HIV-1. However, the N-terminal HA-tagged CT-truncated BST-2 retained little antiviral activity and dramatically differed from its original protein in the cell surface level and intracellular localization. Further, we showed that the replacement of the CT domain with a hydrophobic tag altered BST-2 function possibly by preventing its normal vesicular trafficking. Notably, we demonstrated that a positive charged motif “KRXK” in the conjunctive region between the cytotail and the transmembrane domain which is conserved in primate BST-2 is important for the protein trafficking and the antiviral function. These results suggest that although the CT of BST-2 is not essential for its antiviral activity, the composition of residues in this region may play important roles in its normal trafficking which subsequently affected its function. These observations provide additional implications for the structure-function model of BST-2.     

探讨模拟冷空气降温过程对健康大鼠和高血压大鼠凝血功能的影响

目的:通过模拟冷空气温度变化过程给予健康大鼠和高血压大鼠冷刺激,以此探讨冷空气过程对机体凝血功能的影响。方法:收集张掖市2011年3月一次典型冷空气过程数据,利用气象环境模拟箱模拟其温度变化过程。将24只雄性健康大鼠和24只雄性高血压大鼠分别随机分成最低温组(Tmin组)、Tmin对照组、复温组(Tr组)和Tr对照组。将Tmin组和Tr组大鼠放入气候箱中暴露冷空气温度变化过程。在Tmin和Tr时点分别停止Tmin组大鼠和Tr组大鼠冷空气暴露,并采血以测定其凝血功能指标—凝血四项。结果:与对照组相比,健康大鼠和高血压大鼠活化部分凝血酶原时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)在降温后与对照组相比没有显著差异(P>0.05),但血浆纤维蛋白原(Fbg)含量在高血压大鼠和健康大鼠Tmin组明显高于其对照组(P<0.01)。温度恢复后,其在健康组含量仍然高于其对照组(P<0.05),而在高血压大鼠中没有差异(P>0.05)。同Fbg,反应Fbg的纤维蛋白原时间(Fbg-time)在健康大鼠Tmin和Tr组中短于对照组(P<0.01,),而在高血压大鼠中仅在Tmin组短于对照组(P<0.01)。高血压大鼠血中的Fbg含量和Fbgt明显高于和短于健康大鼠(P<0.01)。结论:①冷空气降温过程能增加机体血中Fbg含量,使凝血功能增强,可能增加心血管疾病危险性;②冷空气刺激对健康大鼠凝血功能影响强于高血压大鼠。     

Structure-activity relationship of 5'-substituted fluoro-neplanocin a analogues as potent inhibitors of S-adenosylhomocysteine hydrolase

Four 5'-substituted fluoro-neplanocin A analogues la-d were designed and synthesized, and the inhibitory activity against SAH was in the following order: NH2 > SH > F, N3, indicating a hydrogen bonding donor is essential for inhibitory activity.     

花灌木种质资源及其遗传多样性研究

综述了国内外关于花灌木种质资源在形态学、细胞学、酶、DNA等水平的多样性研究,探讨了目前花灌木种质资源遗传多样性研究中存在的一些问题和建议。     

Periplaneta Americana extract inhibits osteoclastic differentiation in vitro

ObjectivesPeriplaneta americana extract (PAE) is proven to be promising in treating fever, wound healing, liver fibrosis, and cardiovascular disease. However, the role of PAE in skeletal disorders remains unclear. This study investigated whether PAE regulates osteoclastic differentiation in vitro via the culture using RAW264.7 cells and bone marrow derived macrophages (BMDMs).Materials and MethodsRAW264.7 cells and BMDMs were cultured and induced for osteoclastic differentiation supplementing with different concentrations of PAE (0, 0.1, 1, and 10 mg/mL). Cell counting kit‐8 (CCK‐8) assay was used to detect the cytotoxicity and cell proliferation. TRAP staining, actin ring staining, real‐time quantitative PCR (RT‐qPCR), and bone resorption activity test were performed to detect osteoclastic differentiation. RT‐qPCR and enzyme‐linked immunosorbent assay (ELISA) were conducted to assay the expression and secretion of inflammatory factors. RNA sequencing (RNA‐seq) and western blot analysis were carried out to uncover the underlying mechanism.ResultsCCK‐8 results showed that 10 mg/mL and a lower concentration of PAE did not affect cell proliferation. RT‐qPCR analysis verified that PAE down‐regulated the osteoclastic genes Nfatc1, Ctsk, and Acp5 in macrophages. Moreover, PAE restrained the differentiation, formation, and function of osteoclasts. Besides, RT‐qPCR and ELISA assays showed that PAE decreased inflammatory genes expression and reduced the secretion of inflammatory factors, including IL1β, IL6, and TNFα. Subsequent RNA‐seq analysis identified possible genes and signaling pathways of PAE‐mediated osteoclastogenesis suppression.ConclusionsOur study indicates that PAE has inhibitive effects on osteoclastogenesis and may be a potential therapeutic alternative for bone diseases.

Periplaneta americana extract (PAE), the animal medicine material extracted from the insects Periplaneta americana, is proven to possess a variety of pharmacological functions. However, the role of PAE in skeletal disorders remains unclear. In this study, we found that PAE decreased osteoclast genes expression Nfatc1, Ctsk, and Acp5 in macrophages. Besides, PAE restrained the differentiation, formation, and function of osteoclasts. Moreover, PAE suppressed the LPS‐induced inflammation. Subsequent RNA‐seq analysis identified the signaling pathways of PAE‐mediated osteoclastogenesis suppression. Our study indicated that PAE has inhibitive effects on osteoclastic differentiation and may be a potential therapeutic Chinese medicine for bone diseases.     

传输靶向COX-2 siRNA联合化疗药物对大鼠胃癌细胞生长

目的：通过动物实验探讨传输靶向COX-2 siRNA联合化疗药物对大鼠胃癌细胞生长的抑制作用。方法：24 只健康SD 大鼠 平分为三组,治疗组用COX-2-siRNA转染的胃癌SGC7901 细胞接种,同时进行环磷酰胺、丝裂霉素C 化疗治疗;阴性对照组,用 阴性对照siRNA 转染的胃癌SGC7901 细胞接种,同时进行环磷酰胺、丝裂霉素C 化疗治疗;对照组(n=8),用未经转染的胃癌 SGC7901 细胞接种,不进行化疗治疗;三组转染后都接种了裸鼠。结果：治疗组、阴性对照组及对照组胃癌细胞凋亡率分别为 （22.28± 0.12）%、（1.23± 0.17）%和（1.03± 0.14）%,治疗组与阴性对照组和对照组比较差异都有统计学意义(t=18.152,17.555, P<0.05)。治疗组的抑瘤率为76.7%,阴性对照组和对照组分别为12.8%和6.89%,治疗组的抑瘤率明显高于其他两组(x2=15. 211,13.899,P<0.05)。Western blotting检测结果显示治疗组的COX-2 蛋白表达含量得到了明显抑制。结论：传输靶向COX-2 siRNA和化疗药物的配合应用可有效抑制COX-2 蛋白的表达,从而抑制胃癌细胞的生长,从而起到更好的治疗效果。