Caveolin‐1 down‐regulation is required for Wnt5a‐Frizzled 2 signalling in Ha‐RasV12‐induced cell transformation

Caveolin‐1 (Cav1) is down‐regulated during MK4 (MDCK cells harbouring inducible Ha‐Ras^V12 gene) transformation by Ha‐Ras^V12. Cav1 overexpression abrogates the Ha‐Ras^V12‐driven transformation of MK4 cells; however, the targeted down‐regulation of Cav1 is not sufficient to mimic this transformation. Cav1‐silenced cells, including MK4/shCav1 cells and MDCK/shCav1 cells, showed an increased cell area and discontinuous junction‐related proteins staining. Cellular and mechanical transformations were completed when MDCK/shCav1 cells were treated with medium conditioned by MK4 cells treated with IPTG (MK4+I‐CM) but not with medium conditioned by MK4 cells. Nanoparticle tracking analysis showed that Ha‐Ras^V12‐inducing MK4 cells increased exosome‐like microvesicles release compared with their normal counterparts. The cellular and mechanical transformation activities of MK4+I‐CM were abolished after heat treatment and exosome depletion and were copied by exosomes derived from MK4+I‐CM (MK4+I‐EXs). Wnt5a, a downstream product of Ha‐Ras^V12, was markedly secreted by MK4+I‐CM and MK4+I‐EXs. Suppression of Wnt5a expression and secretion using the porcupine inhibitor C59 or Wnt5a siRNA inhibited the Ha‐Ras^V12‐ and MK4+I‐CM‐induced transformation of MK4 cells and MDCK/shCav1 cells, respectively. Cav1 down‐regulation, either by Ha‐Ras^V12 or targeted shRNA, increased frizzled‐2 (Fzd2) protein levels without affecting its mRNA levels, suggesting a novel role of Cav1 in negatively regulating Fzd2 expression. Additionally, silencing Cav1 facilitated the internalization of MK4+I‐EXs in MDCK cells. These data suggest that Cav1‐dependent repression of Fzd2 and exosome uptake is potentially relevant to its antitransformation activity, which hinders the activation of Ha‐Ras^V12‐Wnt5a‐Stat3 pathway. Altogether, these results suggest that both decreasing Cav1 and increasing exosomal Wnt5a must be implemented during Ha‐Ras^V12‐driven cell transformation.     

烧伤早期伴发脓毒症患者血清PCT、hs-CRP、COR及WBC的动态变化及临床意义

目的:探讨烧伤早期伴发脓毒症患者血清降钙素原(PCT)、超敏C反应蛋白(hs-CRP)、皮质醇(COR)和白细胞计数(WBC)水平并分析其临床价值。方法:回顾性分析2014年12月至2018年1月本院收治的63例烧伤早期患者的临床资料,其中19例患者伴发脓毒症(脓毒症组),44例患者未伴发脓毒症(无脓毒症组),另选取30例于本院进行体检的志愿者作为健康对照组,依据病情严重程度将脓毒症患者分为脓毒症休克组(n=4)、严重脓毒症组(n=7)、一般脓毒症组(n=8),依据预后情况分为预后良好组(n=13)和预后不良组(n=6)。动态检测并对比所有研究对象血清PCT、hs-CRP、COR和WBC水平。结果:无脓毒症组、脓毒症组血清PCT、hs-CRP、COR和WBC水平高于健康对照组,且脓毒症组高于无脓毒症组(P<0.05)。烧伤后7 d、11 d、15 d,脓毒症组血清PCT、hs-CRP、COR和WBC水平均高于无脓毒症组(P<0.05)。脓毒症休克组血清PCT、hs-CRP、COR和WBC水平高于严重脓毒症组、一般脓毒症组,且严重脓毒症组患者高于一般脓毒症组患者(P<0.05)。预后不良组血清PCT、hs-CRP、COR和WBC水平高于预后良好组(P<0.05)。结论:血清PCT、hs-CRP、COR和WBC水平在烧伤早期伴发脓毒症患者中水平异常升高,与病情严重程度有关,上述指标对预后判断有一定的临床参考价值。     

舟山群岛生态系统健康与旅游经济协调发展评价

在界定生态系统健康与旅游经济协调发展定义的基础上,构建了海岛目的地生态系统健康和旅游经济协调发展评价的指标体系,采用改进的TOPSIS法对2000—2012年舟山群岛生态系统健康和海岛旅游业的协调发展状况进行了定量评价,运用障碍度模型对其协调发展的障碍因素进行了分析,并使用Logistic模型对2013—2015年协调发展状态进行了预测。研究表明:(1)2000—2012年,舟山群岛海岛生态系统健康和旅游经济的静态协调度和动态协调度总体均呈持增加趋势,静态协调度由0.6453增加到0.7301,动态协调度由0.6453增加至0.6874;(2)2000—2012年,舟山群岛生态系统健康和旅游经济由初级协调发展型向中级协调发展型演化,其中2000—2007年为初级协调发展型,2008—2012年为中级协调发展型;(3)海洋经济占GDP比重、近海海域环境功能区达标率、环保投入占GDP比重、公路网密度、城镇化率是影响舟山群岛生态系统健康和旅游经济协调发展的主要障碍因子;(4)2013—2015年,舟山群岛生态系统健康和旅游经济的静态协调度预测值为0.8335、0.8442和0.8543,动态协调度的预测值为0.6885、0.6916和0.6947,说明两者的协调发展状态将持续改善。     

Predicting the progress of colon cancer by DNA methylation markers of the p16 gene in feces - Evidence from an animal model

A new noninvasive screening tool for colorectal neoplasia detects epigenetic alterations exhibited by gastrointestinal tumor cells shed into stool. There is insufficient existing data to determine temporal associations between colorectal cancer (CRC) progression and aberrant DNA methylation. To evaluate the feasibility of using fecal DNA methylation status to determine CRC progression, we collected stool samples from 14 male SD rats aged six weeks, and administered subcutaneous injections of either 1,2-dimethylhydrazine or saline weekly. p16 DNA methylation statuses in tumorous and normal colon tissue, and from stool samples were determined using methylation-specific PCR. Additionally, p16 methylation was detected in stool DNA from 85.7% of the CRC rats. The earliest change in p16 methylation status in the DMH-treated group stool samples occurred during week nine; repeatabilities were 57.1% in week 19 (p = 0.070) and 85.7% in week 34 (p = 0.005). A temporal correlation was evidenced between progression of CRC and p16 methylation status, as evidenced by DMH-induced rat feces. Using fecal DNA methylation status to determine colorectal tissue methylation status can reveal CRC progression. Our data suggests that p16 promoter methylation is a feasible epigenetic marker for the detection and may be useful for CRC screening.     

Symptomatic Versus Inapparent Outcome in Repeat Dengue Virus Infections Is Influenced by the Time Interval between Infections and Study Year

Four dengue virus serotypes (DENV1-4) circulate globally, causing more human illness than any other arthropod-borne virus. Dengue can present as a range of clinical manifestations from undifferentiated fever to Dengue Fever to severe, life-threatening syndromes. However, most DENV infections are inapparent. Yet, little is known about determinants of inapparent versus symptomatic DENV infection outcome. Here, we analyzed over 2,000 DENV infections from 2004 to 2011 in a prospective pediatric cohort study in Managua, Nicaragua. Symptomatic cases were captured at the study health center, and paired healthy annual samples were examined on a yearly basis using serological methods to identify inapparent DENV infections. Overall, inapparent and symptomatic DENV infections were equally distributed by sex. The mean age of infection was 1.2 years higher for symptomatic DENV infections as compared to inapparent infections. Although inapparent versus symptomatic outcome did not differ by infection number (first, second or third/post-second DENV infections), substantial variation in the proportion of symptomatic DENV infections among all DENV infections was observed across study years. In participants with repeat DENV infections, the time interval between a first inapparent DENV infection and a second inapparent infection was significantly shorter than the interval between a first inapparent and a second symptomatic infection. This difference was not observed in subsequent infections. This result was confirmed using two different serological techniques that measure total anti-DENV antibodies and serotype-specific neutralizing antibodies, respectively. Taken together, these findings show that, in this study, age, study year and time interval between consecutive DENV infections influence inapparent versus symptomatic infection outcome, while sex and infection number had no significant effect. Moreover, these results suggest that the window of cross-protection induced by a first infection with DENV against a second symptomatic infection is approximately 2 years. These findings are important for modeling dengue epidemics and development of vaccines.     

缺锌和HCO_3~-处理对诸葛菜和油菜有机酸特征的影响

以诸葛菜和油菜为材料,水培环境下设置4个不同的缺锌和碳酸氢根离子胁迫处理,分别为+Zn0(含Zn且不加HCO3-的处理组),+Zn10(含Zn且加10 mmol·L-1HCO3-的处理组),-Zn0(缺Zn且不加HCO3-的处理组)和-Zn10(缺Zn且加10 mmol·L-1HCO3-的处理组),利用离子色谱法分析了4个处理的两种植物幼苗器官(根、茎、叶)及根系分泌物中的有机酸特征。结果表明:(1)高浓度碳酸氢根离子处理显著增加了两种植物器官及根系分泌的有机酸总量,尤其是在缺锌和高浓度碳酸氢根离子双重胁迫下(-Zn10处理),诸葛菜器官和根系分泌的有机酸比油菜更敏感,草酸、柠檬酸和苹果酸是诸葛菜器官和根系分泌物中的优势酸,这三种有机酸的含量分别占其有机酸总量的75%及以上;(2)叶片是两种植物有机酸产生的主要器官,有机酸的含量和分配比例从地上部分(叶和茎)到地下部分(根)减少;(3)两种植物器官和根系分泌物中的有机酸变化趋势一致,叶片中有机酸主要来源于暗呼吸过程和光呼吸过程,其他器官和根系分泌物中的有机酸主要来源于暗呼吸过程;(4)诸葛菜对缺锌和高浓度碳酸氢根离子的适应能力强于油菜,为诸葛菜的喀斯特适生性和低锌和高浓度碳酸氢根离子环境(如喀斯特环境)的生态修复提供了理论依据。     

Gene transfer to pre-hematopoietic and committed hematopoietic precursors in the early mouse Yolk Sac: a comparative study between <Emphasis Type="Italic">in situ</Emphasis> electroporation and retroviral transduction

Background  

Hematopoietic development in vertebrate embryos results from the sequential contribution of two pools of precursors independently generated. While intra-embryonic precursors harbour the features of hematopoietic stem cells (HSC), precursors formed earlier in the yolk sac (YS) display limited differentiation and self-renewal potentials. The mechanisms leading to the generation of the precursors in both sites are still largely unknown, as are the molecular basis underlying their different potential. A possible approach to assess the role of candidate genes is to transfer or modulate their expression/activity in both sites. We thus designed and compared transduction protocols to target either native extra-embryonic precursors, or hematopoietic precursors.     

Response of cells on surface-induced nanopatterns: fibroblasts and mesenchymal progenitor cells

Ultrathin films of a poly(styrene)-block-poly(2-vinylpyrindine) diblock copolymer (PS-b-P2VP) and poly(styrene)-block-poly(4-vinylpyrindine) diblock copolymer (PS-b-P4VP) were used to form surface-induced nanopattern (SINPAT) on mica. Surface interaction controlled microphase separation led to the formation of chemically heterogeneous surface nanopatterns on dry ultrathin films. Two distinct nanopatterned surfaces, namely, wormlike and dotlike patterns, were used to investigate the influence of topography in the nanometer range on cell adhesion, proliferation, and migration. Atomic force microscopy was used to confirm that SINPAT was stable under cell culture conditions. Fibroblasts and mesenchymal progenitor cells were cultured on the nanopatterned surfaces. Phase contrast and confocal laser microscopy showed that fibroblasts and mesenchymal progenitor cells preferred the densely spaced wormlike patterns. Atomic force microscopy showed that the cells remodelled the extracellular matrix differently as they migrate over the two distinctly different nanopatterns.