大花栀子植物挥发物成分测定及其日变化分析

为探究大花栀子植物挥发物成分组成及其一天内早、中、晚的差异,采用热脱附气质联用技术对其进行香气成分的分析。结果表明:全天从其花朵中共鉴定出62种成分,主要为萜烯类、酯类、醇类物质,且不同时间其成分差异显著,如早、中、晚3个时间段,β-蒎烯相对含量分别为1.93%、1.69%、8.81%,顺式-β-罗勒烯分别为28.22%、4.35%、16.47%。3-蒈烯(3.45%)、异丁子香酚(0.21%)等只在早上检出;月桂烯(0.38%)、伞花烃(2.46%)等只在晚上检出;芳樟醇、金合欢烯等在早上和午间两个时间段相对含量较高,而在晚上却未检测出。从植物VOCs角度结合其日变化动态,为大花栀子园林配置及其综合开发利用提供理论依据。     

Identification and expansion of cancer stem cells in tumor tissues and peripheral blood derived from gastric adenocarcinoma patients

Gastric cancer is the fourth most common cancer worldwide, with a high rate of death and low 5-year survival rate. To date, there is a lack of efficient therapeutic protocols for gastric cancer. Recent studies suggest that cancer stem cells (CSCs) are responsible for tumor initiation, invasion, metastasis, and resistance to anticancer therapies. Thus, therapies that target gastric CSCs are attractive. However, CSCs in human gastric adenocarcinoma (GAC) have not been described. Here, we identify CSCs in tumor tissues and peripheral blood from GAC patients. CSCs of human GAC (GCSCs) that are isolated from tumor tissues and peripheral blood of patients carried CD44 and CD54 surface markers, generated tumors that highly resemble the original human tumors when injected into immunodeficient mice, differentiated into gastric epithelial cells in vitro, and self-renewed in vivo and in vitro. Our findings suggest that effective therapeutic protocols must target GCSCs. The capture of GCSCs from the circulation of GAC patients also shows great potential for identification of a critical cell population potentially responsible for tumor metastasis, and provides an effective protocol for early diagnosis and longitudinal monitoring of gastric cancer.     

Studies on selectable marker for genetic engineering of Zymomonas mobilis ZM4 and CP4 strain

尽管质粒和选择标记的使用作为基因工程最基本的一环而为人们所熟知,但对一些特殊菌种(菌株)或研究很少的菌种(菌株)的基因工程操作来说,质粒和选择标记可能仍然是一个并未完全解决的问题,因而需要不断提高认识、不断改进.运动发酵单胞菌Zymomonas mobilis具有突出的产醇性能,但其多种内源质粒和多种抗性的特点,增加了其基因工程操作时质粒和选择标记选用的难度.本研究在测定四个抗生素即Ap、Cm、Tc、Km对典型菌株ZM4、CP4的最低生长抑制浓度的基础上,初步确定了这两个菌株基因工程操作时的四个抗生素使用浓度依次分别为300、100、25、350μg/mL(ZM4)和500、100、25、250 μg/mL(CP4);并进一步通过穿梭载体pZB21、宽宿主载体pBBR1MCS-2和整合载体pBR328-ldh R-cml - ldhL的转化,初步分析和证明了这些选择标记和在相应抗生素浓度下的效果:首先,对每一个选择标记基因来说,前述抗生素浓度是适于携带此选择标记基因的质粒的转化筛选和相应转化子培养的;其次,在前述抗生素浓度下,综合筛选平板阳性率和转化效率、培养物菌体形态异常程度等指标,四个选择标记基因中,以Cm和Tc抗性标记基因效果最好,Km抗性标记基因居中,Ap抗性标记基因最差.这些结果为ZM4、CP4基因工程遗传改造用抗性标记基因、质粒、抗生素的选择及转化系统的完善奠定了基础.     

Multiple end joining mechanisms repair a chromosomal DNA break in fission yeast

Non-homologous end joining (NHEJ) is an important mechanism for repairing DNA double-strand breaks (DSBs). The fission yeast Schizosaccharomyces pombe has a conserved set of NHEJ factors including Ku, DNA ligase IV, Xlf1, and Pol4. Their roles in chromosomal DSB repair have not been directly characterized before. Here we used HO endonuclease to create a specific chromosomal DSB in fission yeast and examined the imprecise end joining events allowing cells to survive the continuous expression of HO. Our analysis showed that cell survival was significantly reduced in mutants defective for Ku, ligase IV, or Xlf1. Using Sanger sequencing and Illumina sequencing, we have characterized in depth the repair junction sequences in HO survivors. In wild type cells the majority of repair events were one-nucleotide insertions dependent on Ku, ligase IV, and Pol4. Our data suggest that fission yeast Pol4 is important for gap filling during NHEJ repair and can extend primers in the absence of terminal base pairing with the templates. In Ku and ligase IV mutants, the survivors mainly resulted from two types of alternative end joining events: one used microhomology flanking the HO site to delete sequences of hundreds to thousands of base pairs, the other rejoined the break using the HO-generated overhangs but also introduced one- or two-nucleotide base substitutions. The chromosomal repair assay we describe here should provide a useful tool for further exploration of the end joining repair mechanisms in fission yeast.     

Identification of cross-linked peptides from complex samples

We have developed pLink, software for data analysis of cross-linked proteins coupled with mass-spectrometry analysis. pLink reliably estimates false discovery rate in cross-link identification and is compatible with multiple homo- or hetero-bifunctional cross-linkers. We validated the program with proteins of known structures, and we further tested it on protein complexes, crude immunoprecipitates and whole-cell lysates. We show that it is a robust tool for protein-structure and protein-protein-interaction studies.     

A Rapid Surface-Enhanced Raman Scattering Method for the Determination of Trace Hg2+ Using Rhodamine 6G-Aggregated Nanosilver as Probe

In pH 6.0 Na₂HPO₄-NaH₂PO₄ buffer solution and in the presence of cetyltrimethyl ammonium bromide, nanosilver particles were aggregated to a stable suspension. Therein, rhodamine 6G (Rh6G) exhibited three strong surface-enhanced Raman scattering (SERS) peaks at 613, 1,363, and 1,510 cm^?1, and their SERS intensities were enhanced when the concentration of Rh6G increased. In the presence of Hg²⁺, the SERS intensity decreased greatly owing to formation of stable Rh6G-HgBr ₄ ^2? ternary association complex molecules as well as its particles. In the optimal condition, the decreased SERS intensity at 613 cm^?1 responds linearly with the concentration of Hg²⁺ over 25–2,000 nmol/L. Thus, a new sensitive SERS method has been proposed for the determination of trace Hg²⁺ in the water sample, with satisfactory results.     

浙江4地桃花水母的rDNA-ITS序列分析

采用PCR和DNA测序技术,对浙江省4个地点桃花水母(Craspedacusta)的核糖体RNA基因内转录间隔区(rDNA-ITS)进行了扩增与测序,并与GenBank中已有的桃花水母ITS区基因序列进行比对分析,计算了它们的遗传距离,利用MEGA 4.1构建了系统发育树。结果显示:这4个地点的桃花水母与索氏桃花水母(Craspedacusta sowerbyi)的ITS区基因相似度极高,同源性都在97%以上,遗传距离保持在0～0.008之间,在进化树中与索氏桃花水母聚为同一支。研究结果表明,这4地的桃花水母都属于索氏桃花水母(C.sowerbyi)。     

海南省公益林生态系统服务功能及其价值评估研究

构建了海南省公益林生态系统服务功能的评价指标体系,依据2009年国家森林资源清查数据、尖峰岭生态定位站连续观测数据、社会公共数据等数据资料,利用影子工程法、替代工程法等方法,首次对海南全省公益林生态服务功能价值(2010年)进行评估。结果显示,海南省公益林生态系统服务功能2010年总价值为1113.60亿元,相当于当年全省生产总值(GDP)2050亿元的54%,单位面积森林提供的服务功能的价值为14.09万元/hm2。由于考虑森林提供负氧离子的功能,8项森林生态系统服务功能价值贡献值之中,净化大气环境>固碳释氧>生物多样性>水源涵养>森林保护>森林游憩>保育土壤>积累营养物质。     

中、重度牙周炎与冠心病患者白细胞介素17 及血脂水平的比较

目的:研究汉族人群中、重度牙周炎与冠心病的相关性并初步探讨白细胞介素17在二者相关性中的可能作用。方法:检测和分析40名健康者(健康组)、40例中、重度牙周炎患者(牙周炎组)、28例冠状动脉粥样硬化性心脏病患者(冠心病组)及47例患冠心病伴中、重度牙周炎的患者(冠心病+牙周炎组)血清白细胞介素17水平、血脂水平(血清低密度脂蛋白、高密度脂蛋白、胆固醇、总胆固醇和甘油三酯)和牙周临床指数(附着丧失、探诊深度和探诊出血)。结果:单因素方差分析结果显示,健康组、牙周炎组、冠心病组及牙周炎+冠心病组的血清白细胞介素17水平分别为(13.01±1.23)、(24.45±2.13)、(59.90±2.23)和(68.87±3.43)ng/L,各组血清白细胞介素17间的差异具有统计学意义(P<0.05),且经协方差分析校正年龄、受教育状况、血压和体重指数后显示,各组血清白细胞介素17水平间的差异仍具有统计学意义(P<0.05)。多元Logistic回归分析结果显示,中、重度牙周炎患者发生冠心病的可能性高于牙周健康者,其发生冠心病的相对风险率比值比为2.416(P=0.039;95%CI:1.126-6.659)。经协方差分析校正年龄、受教育状况、血压和体重指数后,各组血清总胆固醇水平间差异仍具有统计学意义(P=0.018)。结论:严重的牙周感染可能通过改变白细胞介素17水平,影响全身炎症反应和冠心病的发生及发展,可能是冠心病事件的危险因素之一。     

Crucial Involvement of Tumor-Associated Neutrophils in the Regulation of Chronic Colitis-Associated Carcinogenesis in Mice

Ulcerative colitis (UC) is a major form of chronic inflammation that can frequently progress to colon cancer. Several studies have demonstrated massive infiltration of neutrophils and macrophages into the lamina propria and submucosa in the progression of UC-associated colon carcinogenesis. Macrophages contribute to the development of colitis-associated colon cancer (CAC). However, the role of neutrophils is not well understood. To better understand the involvement of tumor-associated neutrophils (TANs) in the regulation of CAC, we used a mouse CAC model produced by administering azoxymethane (AOM), followed by repeated dextran sulfate sodium (DSS) ingestion. This causes severe colonic inflammation and subsequent development of multiple tumors in mice colon. We observed that colorectal mucosal inflammation became increasingly severe with AOM and DSS treatment. Macrophages infiltrated the lamina propria and submucosa, together with a marked increase in neutrophil infiltration. The chemokine CXCL2 increased in the lamina propria and submucosal regions of the colons of the treated mice, together with the infiltration of neutrophils expressing CXCR2, a specific receptor for CXCL2. This process was followed by neoplastic transformation. After AOM and DSS treatment, the mice showed enhanced production of metalloproteinase (MMP)-9 and neutrophil elastase (NE), accompanied by excessive vessel generation and cell proliferation. Moreover, CXCL2 promoted neutrophil recruitment and induced neutrophils to express MMP-9 and NE in vitro. Furthermore, administration of neutrophil-neutralizing antibodies after the last DSS cycle markedly reduced the number and size of tumors and decreased the expression of CXCR2, CXCL2, MMP-9, and NE. These observations indicate a crucial role for TANs in the initiation and progression of CAC and suggest that the CXCL2–CXCR2 axis might be useful in reducing the risk of UC-associated colon cancer.