Application of a luminous intensity variation fluorescent probe for the detection of ferric ions

A luminous intensity variation fluorescent probe (Probe 1) for the detection of ferric ion was developed. The quantitative range of Fe³⁺ content detected was 0–600 μM with the limit of detection at 0.76 μM. Furthermore, after 20 min of Fe³⁺ addition, the intensity of the luminescence of Probe 1 solution gradually decreased with increase in Fe³⁺ concentration. In addition, the B and G values of these images showed a linear relationship with Fe³⁺ concentration (0–500 μM). Probe 1 was successfully used for the rapid determination of Fe³⁺ concentration in real samples. This study demonstrates that Probe 1 is an excellent tool for the rapid determination of Fe³⁺ content in real samples using a smart phone without professional equipment.     

Molecular dynamics simulations provide insights into the origin of gleevec’s selectivity toward human tyrosine kinases

Protein kinases are critical drug targets against cancer. Since the discovery of Gleevec, a specific inhibitor of Abl kinase, the capability of this drug to distinguish between Abl and other tyrosine kinases, such as Src, has been intensely investigated but the origin of Gleevec’s selectivity to Abl against Src is less studied. Here, we performed molecular dynamics (MD) simulations, dynamical cross-correlation matrices (DCCM), dynamical network analysis, and binding free energy calculations to explore Gleevec’s selectivity based on the crystal structures of Abl, Src, and their common ancestors (ANC-AS) and the two constructed mutation systems (AS→Abl and AS→Src). MD simulations revealed that the conformation of the phosphate-binding loop (P-loop) was altered significantly in the AS→Abl system. DCCM results unraveled that mutations increased anticorrelated motions in the AS→Abl system. Community network analysis suggested that the P-loop established special contacts in the AS→Abl system that are devoid in the AS→Src system. The binding free energy calculations unveiled that the affinity of Gleevec to AS→Abl increased to near the Abl level, whereas its affinity to AS→Src decreased to near the Src level. Analysis of individual residue contributions showed that the differences were located mainly at the P-loop. This study is valuable for understanding the sensitivity of Gleevec to human tyrosine kinases.

Communicated by Ramaswamy H. Sarma     

Identification of potent,selective and orally bioavailable phenyl ((R)-3-phenylpyrrolidin-3-yl)sulfone analogues as RORγt inverse agonists

An X-ray crystal structure of one of our previously discovered RORγt inverse agonists bound to the RORγt ligand binding domain revealed that the cyclohexane carboxylic acid group of compound 2 plays a significant role in RORγt binding, forming four hydrogen bonding and ionic interactions with RORγt. SAR studies centered around the cyclohexane carboxylic acid group led to identification of several structurally diverse and more potent compounds, including new carboxylic acid analogues 7 and 20, and cyclic sulfone analogues 34 and 37. Notably, compounds 7 and 20 were found to maintain the desirable pharmacokinetic profile of 2.     

Reprogramming of Meiotic Chromatin Architecture during Spermatogenesis

1. Download high-res image (201KB)
2. Download full-size image

     

Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: application to cellular metabolism and accumulation studies

A simple and sensitive HPLC method was developed to simultaneously determine CPT-11 and its major metabolite SN-38 in culture media and cell lysates. Camptothecin (CPT) was used as internal standard (I.S.). Compounds were eluted with acetonitrile-50 mM disodium hydrogen phosphate buffer containing 10 mM sodium 1-heptane-sulfonate, with the pH adjusted to 3.0 using 85% (w/v) orthophosphoric acid (27/73, v/v) by a Hyperclon ODS (C18) column (200 mm x 4.6 mm i.d.), with detection at excitation and emission wavelengths of 380 and 540 nm, respectively. The average extraction efficiencies were 96.9-108.3% for CPT-11 in culture media and 94.3-107.2% for CPT-11 in cell lysates; and 87.7-106.8% for SN-38 in culture media and 90.1-105.6% for SN-38 in cell lysates. Within- and between-day precision and accuracy varied from 0.1 to 10.3%. The limit of quantitation (precision and accuracy <20%) was 5.0 and 2.0 ng/ml for CPT-11 and 1.0 and 0.5 ng/ml for SN-38 in culture media and cell lysates, respectively. This method was successfully applied to quantitate the cellular accumulation and metabolism of CPT-11 and SN-38 in H4-II-E, a rat hepatoma cell line.     

拉米夫定降低HBV转基因孕鼠血中HBV载量的初步研究

目的:观察HBV转基因小鼠孕期使用拉米夫定对降低母血HBV含量的作用及其对母胎安全性的影响,探讨孕期采用HBV抗病毒治疗,降低宫内感染发生率的可行性。方法:应用可高水平表达HBsAg的HBV转基因小鼠,分别从孕早期(孕第5天)和孕中期(孕第10天)开始给予拉米夫定(100mg/kg,天)直至分娩,对照组给予生理盐水。检测母鼠血中HBsAg、HBV DNA等感染指标的变化,同时从多阶段、多方面,用多指标观察妊娠期,尤其是妊娠早期,使用药物对母鼠、胎鼠及新生鼠安全性的影响。结果:21只孕期接受抗病毒药物的HBV转基因母鼠,无一例发现明显的毒性反应,对胎鼠无致畸作用,且不影响新生鼠早期生长发育,同时小鼠病毒DNA水平明显下降。结论:HBV转基因小鼠孕期服用拉米夫定可降低病毒负荷,有助于减少HBV母婴传播的危险性,且对母鼠及其子代的安全性无明显影响。     

基于线粒体CO Ⅰ基因的闪蛱蝶亚科系统发育关系研究

测定了重要林业害虫闪蛱蝶亚科Apaturinae 11属17种蝶类的线粒体细胞色素氧化酶Ⅰ的部分序列,并结合由GenBank下载的该亚科4种蝶类的相应序列进行分析,探讨了闪蛱蝶亚科各属间的系统发育关系。以玉杵带蛱蝶Athymajina,白斑眼蝶Penthema adelma,忘忧尾蛱蝶Polyura nepenthes和白带螯蛱蝶Charaxes bernardus作为外群,采用PAUP4.0b4a软件构建了闪蛱蝶亚科的MP和NJ分子系统树。虽然COI基因数据中第3位点的转换替换已达饱和,但由于这些位点含有大量系统发育信息,因而在数据统计分析时并没有将这些位点删除。同时通过对各分枝稳定性的比较,研究在简约分析中不同转换/颠换加权方式对假定所有特征具有相同权重的影响。分子系统树显示:NJ和不同转换/颠换加权方式下构建的MP系统树中闪蛱蝶亚科均有4个主要的聚类簇,该亚科系统树中存在许多置信度高且稳定的分枝,同时也存在一些因分枝置信度低且不稳定而使其分类地位不能确定的类群。在所构建的系统树中,由分子数据得到的蛱蝶亚科系统发育关系与传统分类学的基本一致,其中迷蛱蝶属Mimathyma为单系群;累积蛱蝶Lelecella limenitoides为明窗蛱蝶Dilipa fenestra的姐妹群且支持二者关系的置信度很高;支持将白斑迷蛱蝶Mimathyma schrenckii,迷蛱蝶M.chevana,夜迷蛱蝶M.nycteis,栗铠蛱蝶Chitoria subcaerulea,黄带铠蛱蝶C.fasciola,铂铠蛱蝶C.pallas,and银白蛱蝶Helcyra subalba等物种由闪蛱蝶属中移出的修订。     

Effects of blood collection conditions on ovarian cancer serum markers

Background

Evaluating diagnostic and early detection biomarkers requires comparing serum protein concentrations among biosamples ascertained from subjects with and without cancer. Efforts are generally made to standardize blood processing and storage conditions for cases and controls, but blood sample collection conditions cannot be completely controlled. For example, blood samples from cases are often obtained from persons aware of their diagnoses, and collected after fasting or in surgery, whereas blood samples from some controls may be obtained in different conditions, such as a clinic visit. By measuring the effects of differences in collection conditions on three different markers, we investigated the potential of these effects to bias validation studies.

Methodology and Principle Findings

We analyzed serum concentrations of three previously studied putative ovarian cancer serum biomarkers–CA 125, Prolactin and MIF–in healthy women, women with ovarian cancer undergoing gynecologic surgery, women undergoing surgery for benign ovary pathology, and women undergoing surgery with pathologically normal ovaries. For women undergoing surgery, a blood sample was collected either in the clinic 1 to 39 days prior to surgery, or on the day of surgery after anesthesia was administered but prior to the surgical procedure, or both. We found that one marker, prolactin, was dramatically affected by collection conditions, while CA 125 and MIF were unaffected. Prolactin levels were not different between case and control groups after accounting for the conditions of sample collection, suggesting that sample ascertainment could explain some or all of the previously reported results about its potential as a biomarker for ovarian cancer.

Conclusions

Biomarker validation studies should use standardized collection conditions, use multiple control groups, and/or collect samples from cases prior to influence of diagnosis whenever feasible to detect and correct for potential biases associated with sample collection.     

Prevalence and risk factors of overweight and obesity in China

Objective: To examine the prevalence and risk factors of overweight and obesity in China. Research Methods and Procedures: A cross‐sectional survey was conducted in a nationally representative sample of 15,540 Chinese adults in 2000–2001. Body weight, height, and waist circumference were measured by trained observers. Overweight and obesity were defined according to the World Health Organization classification. Central obesity was defined according to guidelines of the International Diabetes Federation. Results: Mean BMI and waist circumference were 23.1 kg/m² and 79.6 cm, respectively, for men and 23.5 kg/m² and 77.2 cm, respectively, for women. The prevalences of overweight and obesity were 24.1% and 2.8% in men and 26.1% and 5.0% in women, respectively. The prevalence of central obesity was 16.1% in men and 37.6% in women. The prevalences of overweight, obesity, and central obesity were higher among residents in northern China compared with their counterparts in southern China and among those in urban areas compared with those in rural areas. Lifestyle factors were the most important risk factors to explain the differences in overweight and central obesity between northern and southern residents. Among women, lifestyle and diet were the most important risk factors to explain the differences between urban and rural residents, whereas socioeconomic status, lifestyle, and diet were all important among men. Discussion: Our study indicates that overweight and obesity have become important public health problems in China. Environmental risk factors may be the main reason for regional differences in the prevalence of overweight and obesity in China.