Characterization of Two-Component System CitB Family in Salmonella Pullorum

Salmonella enterica, serovar Gallinarum, biovar Pullorum, is an avian-specific pathogen which has caused considerable economic losses to the poultry industry worldwide. Two-component systems (TCSs) play an essential role in obtaining nutrients, detecting the presence of neighboring bacteria and regulating the expression of virulence factors. The genome analysis of S. Pullorum strain S06004 suggesting the carriage of 22 pairs of TCSs, which belong to five families named CitB, OmpR, NarL, Chemotaxis and LuxR. In the CitB family, three pairs of TCSs, namely CitA-CitB, DcuS-DcuR and DpiB-DpiA, remain unaddressed in S. Pullorum. To systematically investigate the function of the CitB family in S. Pullorum, four mutants, ΔcitAB (abbreviated as Δcit), ΔdcuSR (Δdcu), ΔdpiBA (Δdpi) and ΔcitABΔdcuSRΔdpiBA (Δ3), were made using the CRISPR/Cas9 system. The results demonstrated that the CitB family did not affect the growth of bacteria, the results of biochemical tests, invasion and proliferation in chicken macrophage HD-11 cells and the expression of fimbrial protein. But the mutants showed thicker biofilm formation, higher resistance to antimicrobial agents, enhanced tolerance to inhibition by egg albumen and increased virulence in chicken embryos. Moreover, the deletion of Dpi TCS was detrimental to survival after exposure to hyperosmotic and oxidative environments, as well as the long-term colonization of the small intestine of chickens. Collectively, we provided new knowledge regarding the possible role of the CitB family involved in the pathogenic processes of S. Pullorum.     

Cyt-C Mediated Mitochondrial Pathway Plays an Important Role in Oocyte Apoptosis in Ricefield Eel (Monopterus albus)

Apoptosis plays a key role in the effective removal of excessive and defective germ cells, which is essential for sequential hermaphroditism and sex change in vertebrates. The ricefield eel, Monopterus albus is a protogynous hermaphroditic fish that undergoes a sequential sex change from female to male. Previous studies have demonstrated that apoptosis is involved in sex change in M. albus. However, the apoptotic signaling pathway is unclear. In the current study, we explored the underlying mechanism of apoptosis during gonadal development and focused on the role of the mitochondrial apoptosis signaling pathway in sex change in M. albus. Flow cytometry was performed to detect apoptosis in gonads at five sexual stages and ovary tissues exposed to hydrogen peroxide (H₂O₂) in vitro. Then the expression patterns of key genes and proteins in the mitochondrial pathway, death receptor pathway and endoplasmic reticulum (ER) pathway were examined. The results showed that the apoptosis rate was significantly increased in the early intersexual stage and then decreased with the natural sex change from female to male. Quantitative real-time PCR revealed that bax, tnfr1, and calpain were mainly expressed in the five stages. ELISA demonstrated that the relative content of cytochrome-c (cyt-c) in the mitochondrial pathway was significantly higher than that of caspase8 and caspase12, with a peak in the early intersexual stage, while the levels of caspase8 and caspase12 peaked in the late intersexual stage. Interestingly, the Pearson’s coefficient between cyt-c and the apoptosis rate was 0.705, which suggests that these factors are closely related during the gonadal development of M. albus. Furthermore, the cyt-c signal was found to be increased in the intersexual stage by immunohistochemistry. After incubation with H₂O₂, the mRNA expression of mitochondrial pathway molecules such as bax, apaf-1, and caspase3 increased in ovary tissues. In conclusion, the present results suggest that the mitochondrial apoptotic pathway may play a more important role than the other apoptotic pathways in sex change in M. albus.     

Physapruin A Enhances DNA Damage and Inhibits DNA Repair to Suppress Oral Cancer Cell Proliferation

The selective antiproliferation to oral cancer cells of Physalis peruviana-derived physapruin A (PHA) is rarely reported. Either drug-induced apoptosis and DNA damage or DNA repair suppression may effectively inhibit cancer cell proliferation. This study examined the selective antiproliferation ability of PHA and explored detailed mechanisms of apoptosis, DNA damage, and repair. During an ATP assay, PHA provided high cytotoxicity to two oral cancer cell lines (CAL 27 and Ca9-22) but no cytotoxicity to two non-malignant oral cells (HGF-1 and SG). This selective antiproliferation of PHA was associated with the selective generation of reactive oxygen species (ROS) in oral cancer cells rather than in non-malignant oral cells, as detected by flow cytometry. Moreover, PHA induced other oxidative stresses in oral cancer cells, such as mitochondrial superoxide generation and mitochondrial membrane potential depletion. PHA also demonstrated selective apoptosis in oral cancer cells rather than non-malignant cells in annexin V/7-aminoactinmycin D and caspase 3/7 activity assays. In flow cytometry and immunofluorescence assays, PHA induced γH2AX expressions and increased the γH2AX foci number of DNA damages in oral cancer cells. In contrast, the mRNA expressions for DNA repair signaling, including homologous recombination (HR) and non-homologous end joining (NHEJ)-associated genes, were inhibited by PHA in oral cancer cells. Moreover, the PHA-induced changes were alleviated by the oxidative stress inhibitor N-acetylcysteine. Therefore, PHA generates selective antiproliferation, oxidative stress, and apoptosis associated with DNA damage induction and DNA repair suppression in oral cancer cells.     

Identification and Functional Analysis of Key Autophosphorylation Residues of Arabidopsis Senescence Associated Receptor-like Kinase

Reversible protein phosphorylation mediated by protein kinases and phosphatases plays important roles in the regulation of leaf senescence. We previously reported that the senescence-associated leucine-rich repeat receptor-like kinase AtSARK autophosphorylates on both serine/threonine and tyrosine residues and functions as a positive regulator of Arabidopsis leaf senescence; the senescence-suppressed protein phosphatase SSPP interacts with and dephosphorylates the cytoplasmic domain of AtSARK, thereby negatively regulating leaf senescence. Here, 27 autophosphorylation residues of AtSARK were revealed by mass spectrometry analysis, and six of them, including two Ser, two Thr, and two Tyr residues, were further found to be important for the biological functions of AtSARK. All site-directed mutations of these six residues that resulted in decreased autophosphorylation level of AtSARK could significantly inhibit AtSARK-induced leaf senescence. In addition, mutations mimicking the dephosphorylation form of Ser384 (S384A) or the phosphorylation form of Tyr413 (Y413E) substantially reduced the interaction between AtSARK and SSPP. All results suggest that autophosphorylation of AtSARK is essential for its functions in promoting leaf senescence. The possible roles of S384 and Y413 residues in fine-tuning the interaction between AtSARK and SSPP are discussed herein.     

基于深度学习的真实尺度运动恢复结构方法

传统的多视图几何方法获取场景结构存在两个问题：一是因图片模糊和低纹理带来的特征点误匹配,从而导致重建精度降低;二是单目相机缺少尺度信息,重建结果只能确定未知的比例因子,无法获取准确的场景结构。针对这些问题本文提出一种基于深度学习的真实尺度运动恢复结构方法。首先使用卷积神经网络获取图片的深度信息;接着为了恢复单目相机的尺度信息,引入惯性传感单元(IMU),将IMU获取的加速度和角速度与ORB-SLAM2获取的相机位姿进行时域和频域上的协同,在频域中获取单目相机的尺度信息;最后将图片的深度图和具有尺度因子的相机位姿进行融合,重建出场景的三维结构。实验表明,使用Depth CNN网络获取的单目图像深度图解决了多层卷积池化操作输出图像分辨率低和缺少重要特征信息的问题,绝对值误差达到了0.192,准确率高达0.959;采用多传感器融合的方法,在频域上获取单目相机的尺度能够达到0.24 m的尺度误差,相比于VIORB方法获取的相机尺度精度更高;重建的三维模型与真实大小具有0.2 m左右的误差,验证了本文方法的有效性。     

强流脉冲电子束表面改性镍基合金的研究

DZ22合金因其优良的高温性能被广泛的应用于高温热端构件。为了进一步提高DZ22合金表面性能,使用强流脉冲电子束对其表面进行轰击。本文主要讨论温度场模拟及微观组织状态。采用abaqus模拟软件对轰击时DZ22镍基合金表面的温度场进行模拟。模拟结果表明,轰击时试样表面在0.68μs时达到轰击时表面的最高温度1629℃,超过DZ22镍基高温合金熔点。利用激光共聚焦显微镜对轰击前后合金表面形貌进行观测,实验结果显示,轰击后试样表面产生熔坑形貌。利用X射线衍射仪对轰击前后合金表面组织结构进行分析,实验表明,轰击使合金表面应力状态与晶粒的生长方向发生改变。结果表明,轰击使得DZ22合金表面出现熔坑形貌且改变其晶粒方向。     

不同人群卫生科普需求及传播方式研究

采用查阅资料、现场观察、定量及定性调查等研究方法,以辽宁省农村居民、沈阳市社区居民及北京市农民建筑工为目标人群,了解到农村居民、城市居民、农民建筑工的卫生科普需求以及卫生科普传播方式的评价与选择存在差异,提出调整卫生科普内容、改进卫生科普传播方式及管理工作的建议.     

塔河油田AT1区块T2a3凝析气藏综合治理研究

砂岩凝析气藏见水后开发效果迅速变差,见水后治理成本高、难度大.塔河油田AT1区块T₂a³凝析气藏存在井网分布不均导致局部采速过高、储层非均质性严重、开发初期采气强度偏大、前期认识存在局限性等问题,通过捕捉生产动态中的异常信号,认为气藏存在底油,并据此形成了以油基堵剂堵水等为代表的改善开发效果的针对性技术手段.通过利用隔夹层和平面非均质性进行剩余油气挖潜,取得了较好的治理效果.     

塔河油田碳酸盐岩油藏定量化注水提高水驱效率现场应用与研究

塔河油田碳酸盐岩油藏具有埋藏深、非均质性强的特点,主要储集空间以溶孔、溶洞和裂缝为主,裂缝为主要渗流通道。随着油田能量的递减,注水做为补充能量最经济有效的方式得到大力推广。根据物质平衡原理,依托酸压曲线、注水指示曲线、能量指示曲线,对注水进行定量化计算,精细注水,明确注采关系,优化注采参数,推动油田提高水驱效率和采收率。