Purification of human plasma haptoglobin by hemoglobin-affinity column chromatography

Haptoglobin (Hp) is an acute-phase protein; its plasma levels increase consistently in response to infection and inflammation. The concentration of human plasma Hp is ranged between 1 and 1.5 mg/ml. Similar to blood type, individual human Hp is classified as Hp 1-1, 2-1, or 2-2. The structural and functional analysis of the Hp, however, has not been studied in detail due to its difficult isolation procedure. Previously, we reported a single step for the purification of porcine Hp. In this study, we established a purification method using a high capacity hemoglobin-affinity column. Briefly, DEAE-purified human hemoglobin was first coupled to Sepharose 4B to prepare an affinity column in a 15-ml bed volume. Following a flow through of human plasma and an extensive wash, the bound material was eluted with a solution of 0.15 M NaCl, pH 11 (adjusted by ammonium), to remove low-affinity bound proteins. The high-affinity bound Hp was then eluted with 0.15 M NaCl containing 5 M urea, pH 11, and collected in tubes containing 100 microl of 1 M Tris buffer, pH 7.0. The biological activity of dialyzed Hp was retained as it formed a complex with hemoglobin on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Using this procedure, approximately 10 mg of Hp 1-1, with homogeneity greater than 96%, was obtained from 15 ml of human plasma. Affinity purified Hp 2-1 or 2-2, however, contained trace amounts of apoA-I with the similar approach. The Hp could be further purified by HPLC using a Superose 12 gel-permeation chromatography, if desired, to achieve 100% purity. All the phenotypes of purified Hp consisted of alpha and beta chains on SDS-PAGE in the presence of a reducing reagent, further confirmed by a Western blot analysis. We conclude that human hemoglobin-affinity column was most suitable for the isolation of Hp 1-1 in large quantities. Whereas, one additional step using a gel-permeation was necessary for that of Hp 2-1 and 2-2.     

啤酒酵母多糖的提纯、化学组成及抗氧化性质鉴定

以啤酒酵母为材料,通过化学抽提法得到水溶性提取物Mal;经薄层层析分析表明：Mal主要含有甘露糖;经检测分析,Mal含有蛋白成分,为蛋白多糖;氨基酸自动分析仪分析表明Mal含有多种氨基酸,包括7种必需氨基酸和9种非必需氨基酸;邻苯三酚自氧化法检测发现多糖能够抑制邻苯三酚自氧化,具有抗氧化能力。     

梭形纽带属在中国新纪录及矢梭形纽带绦虫再描述(绦虫纲,鲤蠢目)(英文)

报道了中国鲤蠢目纽带科绦虫1新纪录属和1新纪录种,即梭形纽带属Atractolytocestus Anthony,1958及该属矢梭形纽带绦虫A.sagittaus(Kulakovskaya et Akhmerov,1965).梭形纽带绦虫与该科中国已记录的纽带属和许氏属绦虫的区别分别在于其卵巢后有卵黄腺存在;卵黄腺在子宫和卵巢侧面连续大量分布,睾丸向后延伸到子宫两侧,头节球状或圆锥状.细颈许氏绦虫Khawia tenuicollis Li,1964的分类学地位在该报道中被重新讨论.     

吉林延边智新盆地大拉子组孢粉组合

本文对产自吉林延边地区智新盆地大拉子组上段的孢子花粉进行了系统研究,共计36属91种,另有疑源类化石1属2种,组成以Classopollis-Cicatricosisporites为代表的孢粉组合。根据组合内一些重要分子的地质时限讨论和与相关组合的对比。其时代应归为早白垩世晚期,根据疑源类和主要孢粉属种可能的母体植物生长环境,推测在早白恶世晚期吉林延边地区智新镇是一个宁静的湖泊,气候温暖,干燥。     

Exploring the expression bar code of SAA variants for gastric cancer detection

We reported an integrated platform to explore serum protein variant pattern in cancer and its utility as a new class of biomarker panel for diagnosis. On the model study of serum amyloid A (SAA), we employed nanoprobe‐based affinity mass spectrometry for enrichment, identification and quantitation of SAA variants from serum of 105 gastric cancer patients in comparison with 54 gastritis patients, 54 controls, and 120 patients from other cancer. The result revealed surprisingly heterogeneous and most comprehensive SAA bar code to date, which comprises 24 SAA variants including SAA1‐ and SAA2‐encoded products, polymorphic isoforms, N‐terminal–truncated forms, and three novel SAA oxidized isotypes, in which the variant‐specific peptide sequence were also confirmed by LC‐MS/MS. A diagnostic model was developed for dimension reduction and computational classification of the 24 SAA‐variant bar code, providing good discrimination (AUC = 0.85 ± 3.2E?3) for differentiating gastric cancer group from gastritis and normal groups (sensitivity, 0.76; specificity, 0.81) and was validated with external validation cohort (sensitivity, 0.71; specificity, 0.74). Our platform not only shed light on the occurrence and modification extent of under‐represented serum protein variants in cancer, but also suggested a new concept of diagnostic platform by serum protein variant profile.     

Study on the amount of binding of anti-tumor metal complexes to different target sites of dGMP using trans-[en(2)Os(eta2-H(2))(CF(3)SO(3))](CF(3)SO(3)) in a competitive mode

In order to investigate the binding sites and the amount of binding of a number of anti-tumor metal complexes (cisplatin, Cp(2)TiCl(2) and (CH(3))(2)SnCl(2)) to target molecule DNA mononucleotides in aqueous solution, a 1H NMR recognition probe, trans-[en(2)Os(eta2-H(2))(CF(3)SO(3))](CF(3)SO(3)), was used in a competitive mode. The minimum percentages of binding of anti-tumor metal complexes to different sites of dGMP were also determined.     

Fibroblast growth factor 21 reverses high‐fat diet‐induced impairment of vascular function via the anti‐oxidative pathway in ApoE knockout mice

Circulating endothelial progenitor cells (EPCs), which function in vascular repair, are the markers of endothelial dysfunction and vascular health. Fibroblast growth factor 21 (FGF21), a liver‐secreted protein, plays a crucial role in glucose homeostasis and lipid metabolism. FGF21 has been reported to attenuate the progression of atherosclerosis, but its impact on EPCs under high oxidative stress conditions remains unclear. In vitro studies showed that the β‐klotho protein was expressed in cultured EPCs and that its expression was upregulated by FGF21 treatment. Hydrogen peroxide (H₂O₂)‐induced oxidative stress impaired EPC function, including cell viability, migration and tube formation. Pretreatment with FGF21 restored the functions of EPCs after the exposure to H₂O₂. Administration of N(ω)‐nitro‐L‐arginine methyl ester (L‐NAME), an inhibitor of nitric oxide synthase, inhibited the effects of FGF21 in alleviating oxidative injury by suppressing endothelial nitric oxide synthase (eNOS). In an in vivo study, the administration of FGF21 significantly reduced total cholesterol (TC) and blood glucose levels in apolipoprotein E (ApoE)‐deficient mice that were fed a high‐fat diet (HFD). Endothelial function, as reflected by acetylcholine‐stimulated aortic relaxation, was improved after FGF21 treatment in ApoE‐deficient mice. Analysis of mRNA levels in the aorta indicated that FGF21 increased the mRNA expression of eNOS and upregulated the expression of the antioxidant genes superoxide dismutase (SOD)1 and SOD2 in ApoE‐deficient mice. These data suggest that FGF21 improves EPC functions via the Akt/eNOS/nitric oxide (NO) pathway and reverses endothelial dysfunction under oxidative stress. Therefore, administration of FGF21 may ameliorate a HFD‐induced vascular injury in ApoE‐deficient mice.     

Role of the APOE ε2/ε3/ε4 Polymorphism in the Development of Primary Open-Angle Glaucoma: Evidence from a Comprehensive Meta-Analysis

Primary open-angle glaucoma (POAG) is one of the leading causes of blindness worldwide. The association between the APOE ε2/ε3/ε4 polymorphism and the risk of POAG has been widely reported, but the results of previous studies remain controversial. To comprehensively evaluate the APOE ɛ2/ɛ3/ε4 polymorphism on the genetic risk for POAG, we performed a systematic review and meta-analysis of previously published studies. The PubMed and Web of Science databases were systematically searched to identify relevant studies. Data were extracted from these studies and odds ratios with corresponding 95% confidence intervals were computed to estimate the strength of the association. Stratified analyses according to ethnicity and sensitivity analyses were also conducted for further confirmation. A total of nine studies were eligible for the meta-analysis, and these studies included data on 1928 POAG cases and 1793 unrelated match controls. The combined results showed that there were no associations between the APOE ε2/ε3/ε4 polymorphism and POAG risk in any of the 10 comparison models. The analysis that was stratified by ethnicity subgroups also failed to reveal a significant association. The sensitivity analysis confirmed the stability and reliability of the findings. There was no risk of publication bias. Our meta-analysis provides strong evidence that the APOE ε2/ε3/ε4 polymorphism is not associated with POAG susceptibility in any populations.     

Moduler l’action du glutamate dans le cerveau: de nouvelles pistes ouvertes grâce aux récepteurs métabotropiques

The rapid transmission of information in the central nervous system is mostly mediated by glutamate synapses. The control of this system quickly appeared as a way to modulate, and perhaps normalize, a number of brain dysfunctions. However, the central role of glutamate receptors involved in this transmission, the ionotropic AMPA and NMDA receptors, appeared as an obstacle to the development of drugs devoid of side effects. The discovery of a second family of glutamate receptors, more than fifteen years ago, offered new possibilities to act on the glutamate system. These receptors, the metabotropic glutamate (mGlu) receptors which are coupled to G-proteins, modulate excitatory synaptic transmission. Eight mGlu receptors have been identified and localized either on the post-synaptic element, where they can regulate the AMPA and NMDA receptor activity, or on the presynaptic element, where they control the release of glutamate or other neurotransmitters. Recent data highlights the therapeutic potential of drugs acting at these receptors for the treatment of a variety of pathologies including anxiety, schizophrenia and Parkinson’s disease.