Association of a novel point mutation in MSH2 gene with familial multiple primary cancers

Background

Multiple primary cancers (MPC) have been identified as two or more cancers without any subordinate relationship that occur either simultaneously or metachronously in the same or different organs of an individual. Lynch syndrome is an autosomal dominant genetic disorder that increases the risk of many types of cancers. Lynch syndrome patients who suffer more than two cancers can also be considered as MPC; patients of this kind provide unique resources to learn how genetic mutation causes MPC in different tissues.

Methods

We performed a whole genome sequencing on blood cells and two tumor samples of a Lynch syndrome patient who was diagnosed with five primary cancers. The mutational landscape of the tumors, including somatic point mutations and copy number alternations, was characterized. We also compared Lynch syndrome with sporadic cancers and proposed a model to illustrate the mutational process by which Lynch syndrome progresses to MPC.

Results

We revealed a novel pathologic mutation on the MSH2 gene (G504 splicing) that associates with Lynch syndrome. Systematical comparison of the mutation landscape revealed that multiple cancers in the proband were evolutionarily independent. Integrative analysis showed that truncating mutations of DNA mismatch repair (MMR) genes were significantly enriched in the patient. A mutation progress model that included germline mutations of MMR genes, double hits of MMR system, mutations in tissue-specific driver genes, and rapid accumulation of additional passenger mutations was proposed to illustrate how MPC occurs in Lynch syndrome patients.

Conclusion

Our findings demonstrate that both germline and somatic alterations are driving forces of carcinogenesis, which may resolve the carcinogenic theory of Lynch syndrome.

     

若尔盖湿地退化过程中土壤水源涵养功能

若尔盖湿地是青藏高原上面积最大的沼泽湿地,也是长江、黄河两大河流的水源区,对区域水循环起重要调节作用。近年来在全球变化及放牧的影响下,若尔盖湿地出现了不同程度的退化。为了查明若尔盖湿地退化过程中水源涵养功能的变化趋势,2009年8月对该区域的沼泽草甸、草原草甸和沙化草甸3个阶段的土壤水源涵养功能进行了调查。结果为:若尔盖湿地由沼泽草甸向草原草甸和沙化草甸的退化过程中,土壤容重显著增加(P<0.01),毛管孔隙度和总孔隙度显著下降(P<0.01),且容重和孔隙度在土壤剖面自然分布规律也发生变化;沼泽草甸的土壤自然含水量、毛管持水量、最小持水量和最大持水量均显著高于草原草甸和沙化草甸(P<0.01);0-100 cm 深度范围内的沼泽草甸土壤的最大持水量(8486.27 t/hm²)显著高于草原草甸(4944.98 t/hm²)和沙化草甸(4637.96 t/hm²)(P<0.01)。土壤持水量与有机质含量、毛管孔隙度和总孔隙度有显著正相关(P<0.01),与土壤容重呈显著负相关(P<0.01),并受植被盖度和泥炭层厚度的影响。研究结果表明,若尔盖湿地退化过程中植被盖度、土壤有机质含量及泥炭层厚度的下降和土壤质地沙化是导致若尔盖湿地水源涵养功能下降的主要原因。     

坡度对农田土壤动物群落结构及多样性的影响

为了研究坡度对土壤动物群落的影响,于2010年3月和9月分别对川中丘陵区坡度为5°、15°、25°的3种农田土壤动物进行了调查。共采集土壤动物11657只,隶属4门11纲21目,弹尾目、蜱螨目、颤蚓目和线虫为优势类群。土壤动物群落的类群数在3月随坡度增加无显著变化(P>0.05),9月则呈波动性上升(P<0.01)。群落密度在3月随坡度增加而显著下降(P<0.01),9月的变化趋势则相反(P<0.05)。群落多样性指数在3、9两月均随坡度增加呈显著波动性变化(P<0.05)。坡度对弹尾目、蜱螨目和线虫等主要类群的密度影响显著(P<0.05),并具季节差异。主成分分析(PCA)结果表明坡度对农田土壤动物的群落结构有明显影响,Sorenson和Morisita-Horn相似性系数进一步表明坡度在3月份主要影响土壤动物群落的类群组成,在9月主要影响优势类群的密度。研究结果表明坡度对土壤动物的群落结构、多样性及主要类群的密度有显著影响,并存在季节差异。     

栖息地荒漠化对草原沙蜥食性的影响

栖息地退化会使环境中蜥蜴的可利用食物资源发生改变。采用剖胃法对内蒙古自治区鄂尔多斯市准格尔旗和达拉特旗地区采集的草原沙蜥样本进行了食性分析。对固定沙丘、半固定沙丘和流动沙丘三类样地中77个实胃样本的652只(套)食物进行鉴定和分析的结果表明,这些食物主要为节肢动物,分为35类,隶属3纲11目32科;也在少部分个体(4.69%)的胃容物中发现少量(1.95%)植物碎片。上述三类栖息地中,草原沙蜥对蚁科、蚜科、瓢虫科、叶蝉科和茧蜂科昆虫均有较大的捕食比例;但相比固定沙区栖息地,草原沙蜥在半固定及流动沙丘中增减的食物种类均比较多,导致草原沙蜥在流动沙区与其他两类样地之间有较大的食物相异性。从固定沙丘、半固定沙丘到流动沙丘,随栖息地荒漠化程度的增加,草原沙蜥捕食食物类群的丰富度逐步减少,辛普森优势度指数相应地下降,但营养生态位宽度、香农威纳多样性指数和皮洛均匀性指数却在流动沙丘栖息地上明显增大。这表明鄂尔多斯地区的草原沙蜥是以捕食昆虫等节肢动物为主、兼食极少量植物的杂食性动物。栖息地荒漠化所致的环境变化显著地影响草原沙蜥的食物组成,使其觅食的食物种类逐步减少。     

Cell cycle-dependent expression of volume-activated chloride currents in nasopharyngeal carcinoma cells

Patch-clamping and cell imageanalysis techniques were used to study the expression of thevolume-activated Cl current,I_Cl(vol), and regulatory volume decrease (RVD)capacity in the cell cycle in nasopharyngeal carcinoma cells (CNE-2Z). Hypotonic challenge caused CNE-2Z cells to swell and activated aCl current with a linear conductance, negligibletime-dependent inactivation, and a reversal potential close to theCl equilibrium potential. The sequence of anionpermeability was I > Br > Cl > gluconate. The Cl channelblockers tamoxifen, 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB),and ATP inhibited I_Cl(vol). Synchronous cultures of cells were obtained by the mitotic shake-off technique and by adouble chemical-block (thymidine and hydroxyurea) technique. Theexpression of I_Cl(vol) was cell cycle dependent,being high in G₁ phase, downregulated in S phase, butincreasing again in M phase. Hypotonic solution activated RVD, whichwas cell cycle dependent and inhibited by the Cl channelblockers NPPB, tamoxifen, and ATP. The expression of I_Cl(vol) was closely correlated with the RVDcapacity in the cell cycle, suggesting a functional relationship.Inhibition of I_Cl(vol) by NPPB (100 µM)arrested cells in G₀/G₁. The data also suggest that expression of I_Cl(vol) and RVD capacity areactively modulated during the cell cycle. The volume-activatedCl current associated with RVD may therefore play animportant role during the cell cycle progress.

     

皂荚皂甙提取工艺的研究

研究了以皂荚为原料,对传统有机溶剂提取与水提取和超临界CO2萃取三萜皂甙进行了比较,得出水最适合提取皂荚皂甙;水提工艺部分,在单因素实验的基础上采用正交设计实验优化工艺,得出提取皂荚皂甙的最佳工艺。     

荷木整树蒸腾对干湿季土壤水分的水力响应

降雨在时间上的非均匀分配导致森林土壤含水量呈现明显的干、湿季变化,并可能在干季形成水分胁迫,引起植物蒸腾变化。在监测环境因子的同时,利用Granier热消散探针连续监测荷木(Schima superba)的树干液流,以液流密度值计算整树蒸腾,并结合水力导度与叶片/土壤的水势差,探讨环境因子和水力导度对荷木整树蒸腾的协同控制。结果表明,华南地区的季节性降雨形成的干、湿季并未引起荷木蒸腾在季节上的显著差异,但对产生蒸腾的水力生理产生了显著影响。荷木蒸腾在干、湿季均与主要驱动环境因子(光合有效辐射PAR和水汽压亏缺VPD)呈显著正相关。在水热充足的湿季,荷木蒸腾主要受气孔导度调节;在干季,当空气水汽压亏缺达2.132 MPa时,水力导度与气孔导度协同控制蒸腾。整树水力导度对整树蒸腾的水力补偿出现在15:00—17:00,平均补偿值为0.08 g/s。利用蒸腾的估测值与实测值之间的差值量化荷木的水力补偿效应,是对水力导度与气孔导度协同控制树木蒸腾机理的深入探索。研究结果对于掌握季节性降雨不均背景下华南地区主要造林树种需水和耗水规律,有效发挥森林保水功能具有重要意义。     

High-level expression of the <Emphasis Type="Italic">Penicillium notatum</Emphasis> glucose oxidase gene in <Emphasis Type="Italic">Pichia pastoris</Emphasis> using codon optimization

The glucose oxidase (GOD) gene from Penicillium notatum was expressed in Pichia pastoris. The 1,815 bp gene, god-w, encodes 604 amino acids. Recombinant GOD-w had optimal activity at 35–40°C and pH 6.2 and was stable, from pH 3 to 7 maintaining >75% maximum activity after incubation at 50°C for 1 h. GOD-w worked as well as commercial GODs to improve bread making. To achieve high-level expression of recombinant GOD in P. pastoris, 272 nucleotides involving 228 residues were mutated, consistent with the codon bias of P. pastoris. The optimized recombinant GOD-m yielded 615 U ml⁻¹ (2.5 g protein l⁻¹) in a 3 l fermentor—410% higher than GOD-w (148 U ml⁻¹), and thus is a low-cost alternative for the bread baking industry.     

The Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor contains an immunoreceptor tyrosine-based inhibitory motif that activates Shp2

The Kaposi's sarcoma-associated herpesvirus (KSHV) G protein-coupled receptor (vGPCR) is a constitutively active, highly angiogenic homologue of the interleukin-8 (IL-8) receptors that signals in part via the cytoplasmic protein tyrosine phosphatase Shp2. We show that vGPCR contains a bona fide immunoreceptor tyrosine-based inhibitory motif (ITIM) that binds and constitutively activates Shp2.     

Glycogen-targeting subunits and glucokinase differentially affect pathways of glycogen metabolism and their regulation in hepatocytes

Overexpression of the glucose-phosphorylating enzyme glucokinase (GK) or members of the family of glycogen-targeting subunits of protein phosphatase-1 increases hepatic glucose disposal and glycogen synthesis. This study was undertaken to evaluate the functional properties of a novel, truncated glycogen-targeting subunit derived from the skeletal muscle isoform G(M)/R(Gl) and to compare pathways of glycogen metabolism and their regulation in cells with overexpressed targeting subunits and GK. When overexpressed in hepatocytes, truncated G(M)/R(Gl) (G(M)DeltaC) was approximately twice as potent as full-length G(M)/R(Gl) in stimulation of glycogen synthesis, but clearly less potent than GK or two other native glycogen-targeting subunits, G(L) and PTG. We also found that cells with overexpressed G(M)DeltaC are unique in that glycogen was efficiently degraded in response to lowering of media glucose concentrations, stimulation with forskolin, or a combination of both maneuvers, whereas cells with overexpressed G(L), PTG, or GK exhibited impairment in one or both of these glycogenolytic signaling pathways. (2)H NMR analysis of purified glycogen revealed that hepatocytes with overexpressed GK synthesized a larger portion of their glycogen from triose phosphates and a smaller portion from tricarboxylic acid cycle intermediates than cells with overexpressed glycogen-targeting subunits. Additional evidence for activation of distinct pathways of glycogen synthesis by GK and targeting subunits is provided by the additive effect of co-overexpression of the two types of proteins upon glycogen synthesis and a much larger stimulation of glucose utilization, glucose transport, and lactate production elicited by GK. We conclude that overexpression of the novel targeting subunit G(M)DeltaC confers unique regulation of glycogen metabolism. Furthermore, targeting subunits and GK stimulate glycogen synthesis by distinct pathways.