Comparison of three common DNA concentration measurement methods

Accurate measurement of DNA concentration is important for DNA-based biological applications. DNA concentration is usually determined by the ultraviolet (UV) absorption, fluorescence staining, and diphenylamine reaction methods. However, the best method for quality assurance of measurements is unknown. Here, we comprehensively compared these methods using different types of samples. We found that all three methods accurately determined the concentrations of high-purity DNA solutions. After digestion of DNA samples, concentration measurements revealed that the PicoGreen dye method was very sensitive to the degradation of DNA. The three methods displayed different anti-jamming ability when contaminants such as transfer RNA (tRNA), protein, and organic chemicals were included in DNA solutions. The diphenylamine reaction method gave the highest accuracy, with an average error of approximately 10% between measured and true values. The PicoGreen dye method was influenced by tRNA and protein, and the UV absorption method was susceptible to all kinds of impurities. Overall, the diphenylamine reaction method gave the most accurate results when DNA was mixed with contaminants, the PicoGreen dye method was most suitable for degraded DNA samples or DNA extracted from processed products, and the UV absorbance method was best for evaluating the impurities in DNA solutions.     

Association between TLR4 (+896A/G and +1196C/T) Polymorphisms and Gastric Cancer Risk: An Updated Meta-Analysis

Background

Toll-like receptor 4 (TLR4) is a receptor of lipopolysaccharide in the signaling transduction of gastric epithelial cell. It plays a pivotal role in activation of innate immunity and pathogen recognition and thus acts as a modulator in the development and progression of gastric cancer. Growing studies explored the association of polymorphisms in TLR4 with susceptibility to gastric cancer, but the results have remained controversial and conflicting. To investigate the effect of two selected TLR4 (+896A/G and +1196C/T) polymorphisms on gastric cancer, we performed a meta-analysis.

Methods

A comprehensive search was conducted to identify all eligible case-control publications investigating the association between TLR4 polymorphisms and gastric cancer risk. Odds ratios (OR) and corresponding 95% confidence intervals (CI) were used to assess such association.

Results

Up to March 26 2014, 10 published case-control studies from PubMed and EMBase were available, involving a total of 1888 gastric cancer patients and 3433 control subjects. In the overall meta-analyses, a significantly increased gastric cancer risk was detected in TLR4 +896A/G polymorphism (heterozygous model, AG vs. AA: OR = 1.67, 95% CI, 1.39–2.01; additive model, G vs. A: OR = 1.64, 95% CI, 1.37–1.95) and TLR4 +1196C/T polymorphism (heterozygous model, CT vs. CC: OR = 1.42, 95% CI, 1.11–1.81; additive model, T vs. C: OR = 1.36, 95% CI, 1.08–1.72), similar results were obtained in the subgroup analyses of Caucasian, whereas no associations were detected in any genetic models of non-Caucasian.

Conclusions

The overall results suggest that TLR4 polymorphisms (+896A/G and +1196C/T) may be associated with a significantly increased gastric cancer risk in Caucasian.     

Association of Plasma IL-6 and Hsp70 with HRV at Different Levels of PAHs Metabolites

Background

Exposure to polycyclic aromatic hydrocarbons (PAHs) is associated with reduced heart rate variability (HRV), a strong predictor of cardiovascular diseases, but the mechanism is not well understood.

Objectives

We hypothesized that PAHs might induce systemic inflammation and stress response, contributing to altered cardiac autonomic function.

Methods

HRV indices were measured using a 3-channel digital Holter monitor in 800 coke oven workers. Plasma levels of interleukin-6 (IL-6) and heat shock protein 70 (Hsp70) were determined using ELISA. Twelve urinary PAHs metabolites (OH-PAHs) were measured by gas chromatography-mass spectrometry.

Results

We found that significant dose-dependent relationships between four urinary OH-PAHs and IL-6 (all P _trend<0.05); and an increase in quartiles of IL-6 was significantly associated with a decrease in total power (TP) and low frequency (LF) (P _trend = 0.014 and 0.006, respectively). In particular, elevated IL-6 was associated in a dose-dependent manner with decreased TP and LF in the high-PAHs metabolites groups (all P _trend<0.05), but not in the low-PAHs metabolites groups. No significant association between Hsp70 and HRV in total population was found after multivariate adjustment. However, increased Hsp70 was significantly associated with elevated standard deviation of NN intervals (SDNN), TP and LF in the low-PAHs metabolites groups (all P _trend<0.05). We also observed that both IL-6 and Hsp70 significantly interacted with multiple PAHs metabolites in relation to HRV.

Conclusions

In coke oven workers, increased IL-6 was associated with a dose-response decreased HRV in the high-PAHs metabolites groups, whereas increase of Hsp70 can result in significant dose-related increase in HRV in the low-PAHs metabolites groups.     

Scintigraphic Detection of Dual Ectopic Thyroid Tissue: Experience of a Chinese Tertiary Hospital

Purpose

To assess scintigraphic pattern, clinical indication and relevance of dual ectopic thyroid tissue (ETT). Literature is reviewed for such cases.

Methods

In this 5-year retrospective study, we reviewed all thyroid scintigraphies in our data base. Patients diagnosed with suspected ETT were identified. Literature is reviewed. Statistics were done by one-way analysis of variance and least significant difference test.

Results

From 11905 thyroid scintigraphies during the 5-year period, we retrieved 121 patients eligible for analysis. The top two indications were assessing a palpable front neck mass to determine whether it was an ETT, and primary hypothyroidism. Patients were divided into 3 groups. Group 1 with single ETT (83 cases); group 2 with dual ETT (6 cases) and group 3 with athyroid (32 cases). Age and thyroid hormones were highest in group 2, and lowest in group 3. Thyrotropin was highest in group 3, and lowest in group 2. Thyroxine was given to hypothyroid patients, while no surgery was performed. There were 42 published cases with dual ETT, most of whom were under 30 years old. 38.10% of them were euthyroid, 33.33% hypothyroid, and 21.43% subclinical hypothyroid. Most frequent ectopic positions included lingual (33.73%), sublingual (27.71%) and subhyoid (22.89%).

Conclusions

In our cohort, incidence of dual ETT was 0.05% if the denominator was total number of thyroid scintigraphies. The incidence was 4.96% if the denominator was the number of patients with suspected ETT. Important clinical indication is a front neck palpable mass suggestive of an ETT. Important clinical relevance of recognizing the dual ETT pattern is to avoid inappropriate surgery. After reviewing all published cases, we find dual ETT is often seen in young patients. Most of such patients are euthyroid or mildly hypothyroid. Thyroid ectopia often resides in lingual, sublingual and subhyoid areas.     

The IL-6–STAT3 axis mediates a reciprocal crosstalk between cancer-derived mesenchymal stem cells and neutrophils to synergistically prompt gastric cancer progression

Emerging evidence indicate that mesenchymal stem cells (MSCs) affect tumor progression by reshaping the tumor microenvironment. Neutrophils are essential component of the tumor microenvironment and are critically involved in cancer progression. Whether the phenotype and function of neutrophils is influenced by MSCs is not well understood. Herein, we investigated the interaction between neutrophils and gastric cancer-derived MSCs (GC-MSCs) and explored the biological role of this interaction. We found that GC-MSCs induced the chemotaxis of neutrophils and protected them from spontaneous apoptosis. Neutrophils were activated by the conditioned medium from GC-MSCs with increased expression of IL-8, TNFα, CCL2, and oncostatin M (OSM). GC-MSCs-primed neutrophils augmented the migration of gastric cancer cells in a cell contact-dependent manner but had minimal effect on gastric cancer cell proliferation. In addition, GC-MSCs-primed neutrophils prompted endothelial cells to form tube-like structure in vitro. We demonstrated that GC-MSCs stimulated the activation of STAT3 and ERK1/2 pathways in neutrophils, which was essential for the functions of activated neutrophils. We further revealed that GC-MSCs-derived IL-6 was responsible for the protection and activation of neutrophils. In turn, GC-MSCs-primed neutrophils induced the differentiation of normal MSCs into cancer-associated fibroblasts (CAFs). Collectively, our results suggest that GC-MSCs regulate the chemotaxis, survival, activation, and function of neutrophils in gastric cancer via an IL-6–STAT3–ERK1/2 signaling cascade. The reciprocal interaction between GC-MSCs and neutrophils presents a novel mechanism for the role of MSCs in remodeling cancer niche and provides a potential target for gastric cancer therapy.Accumulating evidence suggest that neutrophils are critical for cancer initiation and progression.^{1, 2} The increased presence of intratumoral neutrophils has been linked to a poorer prognosis for patients with renal cancer, hepatocellular carcinoma (HCC), melanoma, head and neck squamous cell carcinoma (HNSCC), pancreatic cancer, colorectal carcinoma, and gastric adenocarcinoma.³ Recent studies using murine tumor models or involving cancer patients have suggested an important functional role of neutrophils during tumor progression.^{4, 5, 6, 7} Neutrophils-derived factors promote genetic mutations leading to tumorigenesis or promote tumor cell proliferation,⁸ migration, and invasion.^{9, 10} Neutrophils have also been demonstrated to induce tumor vascularization by the production of pro-angiogenic factors^{11, 12}The infiltration of neutrophils into tumors has been shown to be mediated by factors produced by both tumor and stromal cells. Recent reports suggest that tumor cells actively modulate the functions of neutrophils. Tumor-derived CXCL5 modulates the chemotaxis of neutrophils, which in turn enhances the migration and invasion of human HCC cells.¹³ HNSCC cells-derived MIF induces the recruitment and activation of neutrophils through a p38-dependent manner.^{14, 15} Neutrophils respond to hyaluronan fragments in tumor supernatants via PI3K/Akt signaling, leading to prolonged survival and stimulating effect on HCC cell motility.¹⁶ Kuang et al.¹⁷ suggest that IL-17 promotes the migration of neutrophils into HCC through epithelial cell-derived CXC chemokines, resulting in increased MMP-9 production and angiogenesis at invading tumor edge However, much less is known about the role of stromal cells in modulating the phenotype and function of neutrophils in cancer thus far.Cancer-associated fibroblasts (CAFs) have a key role in cancer mainly through secretion of soluble factors, as growth factors or inflammatory mediators, as well as production of extracellular matrix proteins and their proteases. These activated fibroblasts are involved in creating a niche for cancer cells, promoting their proliferation, motility and chemoresistance. Activated fibroblasts express several mesenchymal markers such as α-smooth muscle actin (α-SMA), fibroblast activation protein (FAP), and vimentin. CAFs actively participate in reciprocal interaction with tumor cells and with other cell types in the microenvironment, contributing to a tumor-permissive niche and promoting tumor progression.Mesenchymal stem cells (MSCs) are adult stromal cells with self-renewal and pluripotent differentiation abilities. MSCs can be mobilized from bone marrow to the site of damage, respond to the local microenvironment, and exert wound repair and tissue regeneration functions upon injury and inflammation conditions.¹⁸ MSCs have been considered as one of the major components of the tumor stroma and are believed to be the precursors of CAFs.^{19, 20} We have previously demonstrated that human bone marrow MSCs prompt tumor growth in vivo.²¹ In addition, we have recently isolated MSCs-like cells from the gastric cancer tissues (GC) and the adjacent normal tissues (GCN) and shown that the gastric cancer-derived MSCs (GC-MSCs) possess the properties of CAFs.^{22, 23} As tumor-derived MSCs are often exposed to distinct inflammatory cells and factors in the tumor microenvironment, they may acquire novel functions that are not present in normal MSCs, and these unique functions may have a role in reshaping the tumor microenvironment and ultimately affect tumor progression. As neutrophils are key mediators of tumor progression and tumor angiogenesis, it is likely that an intense interaction may exist between the tumor-derived MSCs and tumor-infiltrating neutrophils.The emerging roles of CAFs in cancer immunoeditting led us to investigate whether GC-MSCs are able to regulate the phenotype and function of neutrophils in gastric cancer. We have shown that there is a reciprocal interaction between GC-MSCs and neutrophils. GC-MSCs enhanced the chemotaxis of peripheral blood-derived neutrophils and protected them from spontaneous apoptosis. GC-MSCs induced the activation of neutrophils to highly express IL-8, CCL2, TNFα, and oncostatin M (OSM), leading to the increase of gastric cancer cell migration and angiogenesis in vitro. GC-MSCs exerted this effect through the IL-6–STAT3–ERK1/2 signaling axis, and blockade of the IL-6–IL-6R interaction or pharmacological inhibition of STAT3 and ERK1/2 activation abrogated this role. In turn, GC-MSCs-activated neutrophils could trigger the CAF differentiation of normal MSCs. Therefore, these results establish a bi-directional interaction between GC-MSCs and neutrophils that may be critically involved in the progression of gastric cancer.     

龙苍沟国家森林公园7种花楸属植物的叶解剖特征及其环境适应性

采用石蜡切片法对四川省龙苍沟国家森林公园内7种花楸属（Sorbus）植物的叶解剖特征进行研究,探究其结构特征与生境的相关性。结果显示：7种植物的叶片均为典型的背腹叶;叶片厚度介于108.16~208.21 μm,种间差异极显著（P<0.01）;上表皮厚度均大于下表皮厚度,且复叶物种的下表皮细胞均有乳突;栅栏组织由1~2层细胞构成,仅多对西康花楸（S. prattii var. aestivalis）的栅海比（栅栏组织与海绵组织的厚度比）为1.93,其余6种植物的栅海比均小于1;中脉维管束均呈心型,为典型的外韧型维管束,种间中脉突起度存在极显著差异（P<0.01）。各解剖结构中,上、下表皮可塑性最大,在生境中具有较强的潜在适应能力;中脉可塑性最小,整体结构较为稳定。栅栏组织、海绵组织和中脉组织是7种植物中种间差异最大的解剖结构。叶解剖结构与生境因子的相关性分析表明,栅栏组织厚度、栅海比和紧密度与年降水量、最暖季降水和海拔正相关（P<0.05）,与季节性温差负相关（P<0.05）;中脉直径和突起度与季节性温差呈正相关（P<0.01）,与年降水量、最暖季降水和海拔正相关负相关（P<0.05）。叶解剖结构性状的适应性变化,体现了7种花楸属植物在龙苍沟国家森林公园的生存策略。     

中国农田土壤动物长期监测样地科学调查监测的实施方法

我国农田土壤动物面临严峻的多样性丧失问题, 建设监测样地并开展长期监测是解决该问题的重要途径, 但至今国内外仍缺乏农田土壤动物长期监测样地科学调查监测的实施方法。依据BCI 50 ha大型固定样地建设规范, 参照我国已建成的森林和农田土壤动物大型固定样地监测经验, 本文提出了农田土壤动物长期监测样地科学调查监测的实施方法。首先, 需要明确科学问题, 确定科学调查监测应遵守的基本原则。其次, 需要规范长期调查监测涉及的专业术语, 依据研究目的和实际情况选择地点和样地, 参照建设规范和农田特征建立农田土壤动物大型固定样地。第三, 以研究农田土壤动物多样性为核心, 揭示土壤动物在农田生态系统健康和功能中的作用, 有选择性地开展4类27项科学指标的长期监测工作, 要求按照统一的、规范化的工作流程开展野外调查和室内实验。最后, 要科学规范地完成标本的鉴定描述和保存保管, 研发体现农田土壤动物特征的数据库和管理信息系统。希望本文的研究结果能推动我国乃至世界范围的规范化样地建设和标准化网络监测, 为我国农田土壤动物评估与保护提供长期可靠的数据支撑。     

昆虫多样性三十年研究进展

当前, 全球昆虫数量和多样性均处于下降趋势, 而导致这一趋势的原因主要包括人为干扰及气候变化。本文基于森林、草地、农业、水生和土壤生态系统, 以植食性、访花、捕食性、寄生性、食果以及食腐昆虫为重点功能昆虫群, 综述了近三十年来国内外昆虫多样性研究领域的主要进展, 并分析了发展趋势。近年来, 昆虫多样性的研究维度不断拓展, 形态多样性研究不断深入, 系统发生多样性、功能多样性和遗传多样性等研究也显著加强。此外, 昆虫多样性研究的空间尺度也逐步扩大, 大尺度区域性研究甚至全球范围的调查持续增长。昆虫进化历史也被引入多样性格局研究中, 并随着系统发生信息学方法的普及而被整合到生态系统建成和生物多样性形成机制研究中。未来需要加强关键昆虫类群整合分类学研究、功能性状多样性、林冠昆虫多样性、互作网络结构等方向的研究。