CRISPR/Cas9-Mediated Generation of Pathogen-Resistant Tomato against Tomato Yellow Leaf Curl Virus and Powdery Mildew

Tomato is one of the major vegetable crops consumed worldwide. Tomato yellow leaf curl virus (TYLCV) and fungal Oidium sp. are devastating pathogens causing yellow leaf curl disease and powdery mildew. Such viral and fungal pathogens reduce tomato crop yields and cause substantial economic losses every year. Several commercial tomato varieties include Ty-5 (SlPelo) and Mildew resistance locus o 1 (SlMlo1) locus that carries the susceptibility (S-gene) factors for TYLCV and powdery mildew, respectively. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a valuable genome editing tool to develop disease-resistant crop varieties. In this regard, targeting susceptibility factors encoded by the host plant genome instead of the viral genome is a promising approach to achieve pathogen resistance without the need for stable inheritance of CRISPR components. In this study, the CRISPR/Cas9 system was employed to target the SlPelo and SlMlo1 for trait introgression in elite tomato cultivar BN-86 to confer host-mediated immunity against pathogens. SlPelo-knockout lines were successfully generated, carrying the biallelic indel mutations. The pathogen resistance assays in SlPelo mutant lines confirmed the suppressed accumulation of TYLCV and restricted the spread to non-inoculated plant parts. Generated knockout lines for the SlMlo1 showed complete resistance to powdery mildew fungus. Overall, our results demonstrate the efficiency of the CRISPR/Cas9 system to introduce targeted mutagenesis for the rapid development of pathogen-resistant varieties in tomato.     

Increased Histone Acetylation and Decreased Expression of Specific Histone Deacetylases in Ultraviolet-Irradiated and Intrinsically Aged Human Skin In Vivo

Histone deacetylases (HDACs) are conserved enzymes that remove acetyl groups from lysine side chains in histones and other proteins and play a crucial role in epigenetic regulation. Previously, we showed that histone acetylation is implicated in ultraviolet (UV)-induced inflammation and matrix impairment. To elucidate the histone acetylation status and specific HDACs involved in skin aging, we examined the changes in histone acetylation, global HDAC activity, and the expression of HDACs and sirtuins (SIRTs) in intrinsically aged and photoaged human skin as well as in UV-irradiated human skin in vivo. Following acute UV irradiation, the acetylated histone H3 (AcH3) level was increased, but HDAC activity and the expression levels of HDAC4, HDAC11, and SIRT4 were significantly decreased. In intrinsically aged skin, AcH3 levels were increased, but HDAC activity and the expression levels of HDAC4, HDAC5, HDAC10, HDAC11, SIRT6, and SIRT7 were significantly decreased. However, histone acetylation and HDAC expression in photoaged skin were not significantly different from those in intrinsically aged skin. Collectively, HDAC4 and HDAC11 were decreased in both UV-irradiated and intrinsically aged skin, suggesting that they may play a universal role in increased histone acetylation associated with skin aging.     

Identification of Cyclophilin A as a Potential Anticancer Target of Novel Nargenicin A1 Analog in AGS Gastric Cancer Cells

We recently discovered a novel nargenicin A1 analog, 23-demethyl 8,13-deoxynargenicin (compound 9), with potential anti-cancer and anti-angiogenic activities against human gastric adenocarcinoma (AGS) cells. To identify the key molecular targets of compound 9, that are responsible for its biological activities, the changes in proteome expression in AGS cells following compound 9 treatment were analyzed using two-dimensional gel electrophoresis (2-DE), followed by MALDI/TOF/MS. Analyses using chemical proteomics and western blotting revealed that compound 9 treatment significantly suppressed the expression of cyclophilin A (CypA), a member of the immunophilin family. Furthermore, compound 9 downregulated CD147-mediated mitogen-activated protein kinase (MAPK) signaling pathway, including c-Jun N-terminal kinase (JNK) and extracellular signal-regulated protein kinase 1/2 (ERK1/2) by inhibiting the expression of CD147, the cellular receptor of CypA. Notably, the responses of AGS cells to CypA knockdown were significantly correlated with the anticancer and antiangiogenic effects of compound 9. CypA siRNAs reduced the expression of CD147 and phosphorylation of JNK and ERK1/2. In addition, the suppressive effects of CypA siRNAs on proliferation, migration, invasion, and angiogenesis induction of AGS cells were associated with G2/M cell cycle arrest, caspase-mediated apoptosis, inhibition of MMP-9 and MMP-2 expression, inactivation of PI3K/AKT/mTOR pathway, and inhibition of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) expression. The specific interaction between compound 9 and CypA was also confirmed using the drug affinity responsive target stability (DARTS) and cellular thermal shift assay (CETSA) approaches. Moreover, in silico docking analysis revealed that the structure of compound 9 was a good fit for the cyclosporin A binding cavity of CypA. Collectively, these findings provide a novel molecular basis for compound 9-mediated suppression of gastric cancer progression through the targeting of CypA.     

基于IDA方法的地铁车站结构抗震性能评价

针对目前大型地下结构抗震性能评价尚不完善,提出了一种基于非线性增量动力分析(incremental dynamic analysis,IDA)的多层多跨地下结构横断面地震易损性分析的方法.选用典型三层三跨地铁车站建立土-结构二维整体分析有限元模型,采取等效线性化模型模拟土体在地震作用下的剪切模量退化和阻尼增加特性,纤维梁单元模拟多层地下结构在地震作用下的滞回特性.基于PEER强震记录数据库的21条地表地震动峰值加速度中位值,将其统一调幅到不同强度水平,作为非线性IDA的输入地震动.根据IDA结果的统计分析,对于浅埋多层地下结构而言,地表峰值加速度和峰值速度是有效且合适的地震动强度指标,可以用来构建地下结构的易损性曲线.通过与现有的矩形地下结构经验地震易损性曲线和数值地震易损性曲线比较,验证了数值模拟得到的地震易损性曲线的有效性,可以作为快速评价地下结构抗震性能的有效工具.     

走滑断层黏滑错动下隧道破坏的模型试验研究

为了探究断层错动对隧道及围岩的损伤机理及破坏特性,以滇中引水工程香炉山隧洞为依托,通过开展隧道穿越断层破碎带的黏滑错动模型试验,并采用ABAQUS进行数值建模,对试验结果进行验证分析.研究结果表明:断层错动过程中,衬砌主要发生受拉破坏,局部位置衬砌受压破坏;衬砌发生变形及破坏的区域,主要集中在破碎带范围内,以错动面处衬砌变形、破坏最为突出,衬砌的变形随距错动面距离的增大而减少,衬砌横断面内拱顶、右腰位置破坏最为严重;断层错动起始时刻,破碎带中间部位拱顶处衬砌拉损伤已经达到较高量级,衬砌局部开裂破坏,随着断层错动量的增加,损伤不断累积,当错动量达25 mm(相当于实际错动量1 m)时,衬砌整体损伤已累积到较高量级,此时衬砌的破坏接近试验的最终状况.岩体的破裂区域主要集中在距错动面两侧30 cm范围内,围岩变形随着距错动面距离的增加而逐渐减小.     

医院门诊就诊流程优化研究

以业务流程再造、程序分析等相关理论方法为基础,以J市R医院消化内科为例,对医院门诊就诊流程进行优化研究。通过实地走访与问卷调查发现消化内科现行门诊服务流程中存在的问题,基于业务流程再造理论优化门诊服务流程,使用程序分析方法和Flexsim仿真对门诊就诊流程进行对比分析,验证了优化方案的有效性。     

The Role of Chronic Inflammation in Various Diseases and Anti-inflammatory Therapies Containing Natural Products

Chronic inflammation represents a long-term reaction of the body's immune system to noxious stimuli. Such a sustained inflammatory response sometimes results in lasting damage to healthy tissues and organs. In fact, chronic inflammation is implicated in the development and progression of various diseases, including cardiovascular diseases, respiratory diseases, metabolic diseases, neurodegenerative diseases, and even cancers. Targeting nonresolving inflammation thus provides new opportunities for treating relevant diseases. In this review, we will go over several chronic inflammation-associated diseases first with emphasis on the role of inflammation in their pathogenesis. Then, we will summarize a number of natural products that exhibit therapeutic effects against those diseases by acting on different markers in the inflammatory response. We envision that natural products will remain a rich resource for the discovery of new drugs treating diseases associated with chronic inflammation.     

相对压强对白炭黑氮吸附表面积测试稳定性的影响

研究相对压强对白炭黑氮吸附表面积测试稳定性的影响。结果表明:在相同液氮纯度、脱气温度、脱气时间、氮吸附点数量条件下,与相对压强范围为0.05~0.1时相比,相对压强范围为0.05~0.2时白炭黑的氮吸附表面积测量值偏低,且差值随氮吸附表面积的增大呈增大趋势;相对压强范围较小时,氮吸附表面积测试结果的线性相关因数较大,再现性较好,测试结果较稳定。     

陕北±800 kV换流站高填方湿陷性黄土地基处理方案研究

陕北±800 kV换流站用地为原循环工业经济园区挖黄土梁卯、填深沟给出的一块台阶式场地,场地覆盖一层厚度0~50 m的素填土,密实度差异较大,土质极不均匀,填土完成约5年半时间,是一种欠固结土,在其自身重度和大气降水的入渗作用下有自行密实的特点,影响位于填方区建(构)筑物地基的承载力和稳定性。基于该工程特殊的岩土地质情况,除挖方区采用天然地基外,从处理填方区大面积欠固结土和消除部分地基土湿陷性、提高地基土承载力、减小场地沉降变形的角度,对不同地基处理及桩基方案进行技术经济综合比选,提出适合本工程的经济合理的地基处理方案。