北沙参中佛手柑内酯的分离鉴定及体外抗肿瘤活性的初步测定

北沙参为传统的中药材,来源于伞形科(Umbelliferae)植物珊瑚菜(Glehnia littoralis Fr. Schmidt ex Miq. )的干燥根,具有养阴清肺、益胃生津的功效[1],其道地产地为山东莱阳.     

Construct optimization for protein NMR structure analysis using amide hydrogen/deuterium exchange mass spectrometry

Disordered or unstructured regions of proteins, while often very important biologically, can pose significant challenges for resonance assignment and three‐dimensional structure determination of the ordered regions of proteins by NMR methods. In this article, we demonstrate the application of ¹H/²H exchange mass spectrometry (DXMS) for the rapid identification of disordered segments of proteins and design of protein constructs that are more suitable for structural analysis by NMR. In this benchmark study, DXMS is applied to five NMR protein targets chosen from the Northeast Structural Genomics project. These data were then used to design optimized constructs for three partially disordered proteins. Truncated proteins obtained by deletion of disordered N‐ and C‐terminal tails were evaluated using ¹H‐¹⁵N HSQC and ¹H‐¹⁵N heteronuclear NOE NMR experiments to assess their structural integrity. These constructs provide significantly improved NMR spectra, with minimal structural perturbations to the ordered regions of the protein structure. As a representative example, we compare the solution structures of the full length and DXMS‐based truncated construct for a 77‐residue partially disordered DUF896 family protein YnzC from Bacillus subtilis, where deletion of the disordered residues (ca. 40% of the protein) does not affect the native structure. In addition, we demonstrate that throughput of the DXMS process can be increased by analyzing mixtures of up to four proteins without reducing the sequence coverage for each protein. Our results demonstrate that DXMS can serve as a central component of a process for optimizing protein constructs for NMR structure determination. Proteins 2009. © 2009 Wiley‐Liss, Inc.     

LC-MS/MS determination of helicid in human plasma and its application in pharmacokinetic studies

Helicid is a traditional Chinese medicine used to treat headache and insomnia with definite effects. To facilitate pharmacokinetic studies of helicid in man, a sensitive and specific LC-MS/MS method for the quantitative detection of helicid in human plasma was developed and validated. The method involved the addition of bergeninum as the internal standard (IS), protein precipitation, HPLC separation, and quantification by MS/MS system using negative electrospray ionization in the multiple reaction monitoring mode (MRM). The precursor→product ion transitions were monitored at m/z 282.8→120.9 for helicid and m/z 326.9→192.2 for the IS, respectively. The lower limit of quantification (LLOQ) was 0.2 μg/L. The calibration curves for helicid was linear over a concentration range of 0.2-20 μg/L. The intra- and inter-batch analyses of QC samples at 0.4, 2, 20 μg/L indicated good precision (%R.S.D. between 2.69 and 5.47%) and accuracy (between 96.15 and 105.05%). The helicid was stable in human plasma stored at room temperature for at least 24h, 4°C for at least 24h, -20°C for at least 1 month, and for routine three freeze-thaw cycles. This accurate and specific assay provides a useful method for evaluating the pharmacokinetic profile of helicid in humans.     

Seroprevalence of pandemic (H1N1) 2009 in pregnant women in China: an observational study

Background

We investigated the seropositive rates and persistence of antibody against pandemic (H1N1) 2009 virus (pH1N1) in pregnant women and voluntary blood donors after the second wave of the pandemic in Nanjing, China.

Methodology/Principal Findings

Serum samples of unvaccinated pregnant women (n = 720) and voluntary blood donors (n = 320) were collected after the second wave of 2009 pandemic in Nanjing. All samples were tested against pH1N1 strain (A/California/7/2009) with hemagglutination inhibition assay. A significant decline in seropositive rates, from above 50% to about 20%, was observed in pregnant women and voluntary blood donors fifteen weeks after the second wave of the pandemic. A quarter of the samples were tested against a seasonal H1N1 strain (A/Brisbane/59/2007). The antibody titers against pH1N1 strain were found to correlate positively with those against seasonal H1N1 strain. The correlation was modest but statistically significant.

Conclusions and Significance

The high seropositive rates in both pregnant women and voluntary blood donors suggested that the pH1N1 virus had widely spread in these two populations. Immunity derived from natural infection seemed not to be persistent well.     

CCCTC-结合因子与输入蛋白13存在蛋白间相互作用

目的:研究CCCTC-结合因子(CTCF)是否与输入蛋白13(IPO13)存在相互作用。方法:采用GST pull-down及免疫共沉淀实验研究二者的相互作用。结果与结论:实验表明CTCF与IPO13之间存在相互作用     

乌司他丁联合连续性血液净化治疗重症脓毒症临床研究

目的：研究连续性血液净化联合乌司他丁对重症脓毒症患者炎症反应的影响及其临床疗效。方法：70例重症脓毒症患者随机分为对照组（n=22例）、CBP组（n=23例）和CBP＋乌司他丁组（n=25例）,其中对照组采用经典治疗方案,CBP组在此基础上加用连续性血液净化,CBP＋乌司他丁组在CBP组基础上加用乌司他丁治疗。观察比较患者病情发展,分别于治疗前后进行血液生化指标、凝血功能检测和动脉血气分析,ELISA法检测血清CRP水平。结果：①与对照组和CBP组相比,CBP＋鸟司他丁纽患者病死率、ICU住院时间、MODS发生率均明显下降（P〈0．05）。②经过治疗,CBP＋乌司他丁组患者APACHEⅡ评分降至15．46±3．96,与对照组（18．06±4．25）和CBP组（17．14±5．55）比较差异有统计学意义（P〈0．05）。③治疗后,患者BUN、HR降低程度依次为CBP＋乌司他丁组〉CBP组〉对照组,组问比较差异有显著性（P〈0．05）,而治疗前后PH值、HCO3-、MAP比较差并不明显（P〉0．05）。④CBP＋乌司他丁纽血清CRP含量下降,WBC数量减少,其变化程度明显大于CBP组和对照组（P〈0．05）。⑤对照组、CBP组和CBP＋乌司他丁组患者PT、TT和APTT时间延长,血小板数量下降,其中CBP＋鸟司他丁组PT、APTT时间短于CBP组和对照组（P〈0．05）。结论：连续性血液净化联合乌司他丁可有效抑制脓毒症患者炎症反应,缓解病情,改善患者预后。     

单鼻孔-蝶窦入路垂体瘤切除术的体会

目的：探讨经单鼻孔-蝶窦入路垂体瘤切除手术的效果。方法：应用单鼻孔-蝶窦入路切除22例垂体瘤患者。结果：22例患者手术均顺利进行并于术后3月均获随访,全切除14例,次全切除5例,部分切除3例。结论：经单鼻孔-蝶窦入路显微切除垂体瘤属于微侵袭及功能保护性手术,符合现代神经外科的发展方向,严格把握适应症及禁忌症,完善手术技巧是提高手术效果的关键。     

Heat-Stable Molecule Derived from Streptococcus cristatus Induces APOBEC3 Expression and Inhibits HIV-1 Replication

Although most human immunodeficiency virus type 1 (HIV-1) cases worldwide are transmitted through mucosal surfaces, transmission through the oral mucosal surface is a rare event. More than 700 bacterial species have been detected in the oral cavity. Despite great efforts to discover oral inhibitors of HIV, little information is available concerning the anti-HIV activity of oral bacterial components. Here we show that a molecule from an oral commensal bacterium, Streptococcus cristatus CC5A can induce expression of APOBEC3G (A3G) and APOBEC3F (A3F) and inhibit HIV-1 replication in THP-1 cells. We show by qRT-PCR that expression levels of A3G and A3F increase in a dose-dependent manner in the presence of a CC5A extract, as does A3G protein levels by Western blot assay. In addition, when the human monocytic cell line THP-1 was treated with CC5A extract, the replication of HIV-1 IIIB was significantly suppressed compared with IIIB replication in untreated THP-1 cells. Knock down of A3G expression in THP-1 cells compromised the ability of CC5A to inhibit HIV-1 IIIB infectivity. Furthermore, SupT1 cells infected with virus produced from CC5A extract-treated THP-1 cells replicated virus with a higher G to A hypermutation rate (a known consequence of A3G activity) than virus used from untreated THP-1 cells. This suggests that S. cristatus CC5A contains a molecule that induces A3G/F expression and thereby inhibits HIV replication. These findings might lead to the discovery of a novel anti-HIV/AIDS therapeutic.