胃癌组织长链非编码RNA ZDHHC8P1、MEG3、TUG1表达与临床病理特征及预后的关系研究

目的:探讨胃癌组织长链非编码RNA(lncRNA)DHHC型锌指蛋白8假基因1(ZDHHC8P1)、母系表达基因3(MEG3)、牛磺酸上调基因1(TUG1)表达与临床病理特征和预后的关系。方法:选取2013年1月至2016年1月我院病理科收集的83例胃癌患者经手术切除或胃镜活检的癌组织及癌旁组织石蜡标本,检测胃癌和癌旁组织中ZDHHC8P1、MEG3、TUG1表达。分析ZDHHC8P1、MEG3、TUG1表达与胃癌临床病理特征的关系。随访所有患者,分析ZDHHC8P1、MEG3、TUG1表达与患者预后的关系。结果:胃癌组织中ZDHHC8P1、TUG1表达量均高于癌旁组织(P0.05),MEG3表达量低于癌旁组织(P0.05)。ZDHHC8P1表达与肿瘤直径、分化程度、浸润深度、TNM分期、淋巴结转移、远处转移有关(P0.05),MEG3、TUG1表达与分化程度、浸润深度、淋巴结转移有关(P0.05)。Kaplan-Meier生存曲线分析结果显示ZDHHC8P1、TUG1高表达患者无疾病进展生存(PFS)率、总生存(OS)率低于ZDHHC8P1、TUG1低表达患者(P0.05),MEG3低表达患者PFS、OS率低于MEG3高表达患者(P0.05)。Cox风险回归分析结果显示淋巴结转移、ZDHHC8P1高表达、TUG1高表达、MEG3低表达是胃癌患者不良预后的危险因素(HR=1.613、1.956、2.512、-0.824,P0.05)。结论:胃癌组织中ZDHHC8P1、TUG1呈高表达,MEG3呈低表达,ZDHHC8P1、TUG1、MEG3表达均与胃癌临床病理特征和预后有关,可作为胃癌患者预后评估的辅助指标。     

多重耐药鲍曼不动杆菌的耐药性及其耐药基因分析

目的:探讨多重耐药鲍曼不动杆菌(MDR-Ab)的耐药性及其耐药基因,为临床合理选择抗菌药物提供依据。方法:回顾性分析2018年1月至2018年12月鲍曼不动杆菌感染的住院患者信息。使用VITEK-32微生物分析仪/梅里埃药敏卡片GN13鉴定MDR-Ab 95株。采用聚合酶链式反应(多重PCR)检测MDR-Ab携带相关耐药基因。结果:95株MDR-Ab对头孢类抗菌药物耐药率为100%。对氨苄西林-舒巴坦和头孢哌酮-舒巴坦耐药率分别为95.79%和81.05%,对美罗培南和亚胺培南耐药率分别为56.84%和57.89%,对庆大霉素和阿米卡星耐药率均为88.42%,对环丙沙星和左氧氟沙星耐药率分别为100%和88.42%,对四环素、米诺环素、替加环素耐药率分别为87.37%、16.84%和9.47%,对多粘菌素B耐药率为1.05%。95株MDR-Ab中携带β-内酰胺酶中A类酶耐药基因TEM、PER分别95株和25株,D类酶耐药基因OXA-51、carO和adeB各95株,OXA-23基因90株。携带消毒剂耐药基因qacE 60株。携带16S r RNA甲基化酶耐药基因armA 75株。每株MDR-Ab除携带TEM+carO+adeB+OXA-51四种基因外,另同时携带四种基因20株(21.05%),三种基因38株(40.00%)。结论:MDR-Ab对多种抗菌药物的耐药率较高,携带的耐药基因型主要为TEM、carO、adeB及OXA-51。携带多种耐药基因是MDR-Ab耐药重要原因。加强医院感染防控、合理应用抗菌药物对于延缓泛鲍曼不动杆菌耐药性发展具有重要的临床意义。     

人PINK1-shRNA载体的构建及对神经母细胞瘤细胞线粒体的影响

摘要 目的：构建筛选人源靶向特异PINK1-shRNA敲减质粒,转染神经母细胞瘤SH-SY5Y细胞并验证该质粒转染后对PINK1基因的敲减效率,观察对细胞线粒体形态的影响。方法：构建两对人源PINK1-shRNA序列(编号分别为PINK1-shRNA-39和PINK1-shRNA-42),将这2对干扰序列连接在载体上形成重组载体,经测序验证后,将空载体和两对敲减质粒分别转染SH-SY5Y细胞,获得基因敲减的细胞模型,用CCK-8法检测细胞的存活率,用荧光定量PCR方法确定PINK1基因的敲减效率,用蛋白质免疫印迹法验证细胞内PINK1的表达水平是否发生改变,用激光共聚焦显微镜观察线粒体的形态是否发生了改变。结果：我们提取的质粒,经测序结果显示,质粒载体构建成功;转染细胞后,CCK-8 法检测细胞存活率发生了降低,与正常组比较,PINK1-shRNA-39和PINK1-shRNA-42敲减质粒组,细胞的存活率分别降低了13.7%（P＜0.05）和14.1%（P＜0.05）;荧光定量PCR结果显示,与正常组相比,PINK1-shRNA-39组和PINK1-shRNA-42组敲减质粒转染的细胞内PINK1基因的表达分别降低了24.1%（P＜0.01）和36.7%（P＜0.01）;蛋白质印迹法结果显示,与正常组相比,两对质粒分别转染细胞后,PINK1-shRNA-39和PINK1-shRNA-42敲减质粒转染的细胞内PINK1蛋白的表达水平降低,差异有统计学意义(P＜0.01),PINK1-shRNA-42敲减质粒转染的细胞内PINK1蛋白的表达水平有效降低更明显;与正常组比较,激光共聚焦显微镜观察到基因敲减两组细胞的线粒体部分发生断裂,碎片较多,基因敲减两组的线粒体的形态因子显著降低,差异有统计学意义（P＜ 0.01）。结论：成功构建了人源的PINK1基因敲减的质粒,并将敲减的质粒成功转染至SH-SY5Y 细胞中,细胞内PINK1基因的mRNA和蛋白的表达水平降低,且细胞线粒体的形态发生了改变。     

水稻根系遗传育种研究进展

根系作为水稻(Oryza sativa)植株的重要组成部分, 在水稻生长发育过程中发挥多种作用, 包括植物的固定、水分和营养物质的获取以及氨基酸和激素的生物合成等, 其形态结构和生理功能与水稻产量和稻米品质以及抗性等密切相关。目前, 通过遗传及生化等诸多手段, 已挖掘到较多水稻根系QTLs与控制基因。该文综述了水稻根系QTL和基因的研究进展, 并对未来根系研究进行展望, 以期为进一步克隆水稻根系基因和完善水稻理想株型模型提供参考。     

Estimating the potential for coral adaptation to global warming across the Indo‐West Pacific

The potential of reef‐building corals to adapt to increasing sea‐surface temperatures is often debated but has rarely been comprehensively modeled on a region‐wide scale. We used individual‐based simulations to model adaptation to warming in a coral metapopulation comprising 680 reefs and representing the whole of the Central Indo‐West Pacific. Encouragingly, some reefs—most notably Vietnam, Japan, Taiwan, New Caledonia and the southern half of the Great Barrier Reef—exhibited high capacity for adaptation and, in our model, maintained coral cover even under a rapid “business‐as‐usual” warming scenario throughout the modeled period (200 years). Higher resilience of these reefs was observed under all tested parameter settings except the models prohibiting selection and/or migration during warming. At the same time, the majority of reefs in the region tended to collapse within the first 100 years of warming. The adaptive potential (odds of maintaining high coral cover) of a given reef could be predicted based on two metrics: the reef's present‐day temperature, and the proportion of recruits immigrating from warmer locations. The latter metric explains the most variation in adaptive potential, and significantly correlates with actual coral cover changes observed throughout the region between the 1970s and the early 2000s. These findings will help prioritize coral conservation efforts and plan assisted gene flow interventions to boost the adaptive potential of specific coral populations.     

Impact of single nucleotide polymorphisms in the OGG1 and XRCC1 genes on modulation of DNA damage in pesticide-exposed agricultural workers in Punjab,North-West India

Abstract

Pesticide-induced DNA damage is primarily repaired by base excision repair (BER) pathway. However, polymorphism in DNA repair genes may modulate individual’s DNA repair capacity (DRC) leading to increased genotoxicity and adverse health effects. Our first study in North-West Indian population aimed to evaluate the impact of OGG1 rs1052133 (Ser326Cys; C1245G), XRCC1 rs1799782 (Arg194Trp; C26304T) and XRCC1 rs25487 (Arg399Gln; G28152A) polymorphisms on the modulation of pesticide-induced DNA damage in a total of 450 subjects (225 pesticide-exposed agricultural workers and 225 age- and sex-matched controls). DNA damage was estimated by alkaline comet assay using silver-staining method. Genotyping was carried out by PCR-RFLP using site-specific restriction enzymes. Mann-Whitney U-test revealed elevation in DNA damage parameters (p?<?0.01) in pesticide-exposed agricultural workers than controls. Chi-square test showed significant (p?<?0.05) differences in the XRCC1 Arg194Trp (C26304T) and Arg399Gln (G28152A) genotypes among two groups. Multivariate logistic-regression analysis revealed that heterozygous genotypes of OGG1 rs1052133 (326Ser/Cys; 1245CA), XRCC1 rs1799782 (194Arg/Trp; 26304CT) and XRCC1 rs25487 (399Arg/Gln; 2815GA) were positively associated (p?<?0.05) with elevated DNA damage parameters in pesticide-exposed agricultural workers. Our results strongly indicate significant positive association of variant OGG1 and XRCC1 genotypes with reduced DRC and higher pesticide-induced DNA damage in North-West Indian agricultural workers.     

Three proline rotamases involved in calcium homeostasis play differential roles in stress tolerance,virulence and calcineurin regulation of Beauveria bassiana

FK506‐sensitive proline rotamases (FPRs), also known as FK506‐binding proteins (FKBPs), can mediate immunosuppressive drug resistance in budding yeast but their physiological roles in filamentous fungi remain opaque. Here, we report that three FPRs (cytosolic/nuclear 12.15‐kD Fpr1, membrane‐associated 14.78‐kD Fpr2 and nuclear 50.43‐kD Fpr3) are all equally essential for cellular Ca²⁺ homeostasis and contribute significantly to calcineurin activity at different levels in the insect‐pathogenic fungus Beauveria bassiana although the deletion of fpr1 alone conferred resistance to FK506. Radial growth, conidiation, conidial viability and virulence were less compromised in the absence of fpr1 or fpr2 than in the absence of fpr3, which abolished almost all growth on scant media and reduced growth moderately on rich media. The Δfpr3 mutant was more sensitive to Na⁺, K⁺, Mn²⁺, Ca²⁺, Cu²⁺, metal chelate, heat shock and UVB irradiation than was Δfpr2 while both mutants were equally sensitive to Zn²⁺, Mg²⁺, Fe²⁺, H₂O₂ and cell wall‐perturbing agents. In contrast, the Δfpr1 mutant was less sensitive to fewer stress cues. Most of 32 examined genes involved in DNA damage repair, Na⁺/K⁺ detoxification or osmotolerance and Ca²⁺ homeostasis were downregulated sharply in Δfpr2 and Δfpr3 but rarely so affected in Δfpr1, coinciding well with their phenotypic changes. These findings uncover important, but differential, roles of three FPRs in the fungal adaptation to insect host and environment and provide novel insight into their essential roles in calcium signalling pathway.