高效液相色谱法测定红细胞内α-生育酚

目的：寻找一种测定红细胞内α-生育酚含量的较好方法。方法：将含有生育酚的肝素抗凝血离心后吸出血浆,剩余的红细胞用生理盐水洗涤后用氢氧化钾皂化,并加入联苯三酚以防止α-生育酚被氧化。再用正已烷提取红细胞内的α-生育酚,并进行高效液相层析。结果：α-生育酚浓度为0．927～9．267 μmol／L,线性关系良好,最低检测限为0．323μmol／L,变异系数1．3％,回收率100．2％。结论：该方法简单可靠,重复性好,并可从细胞水平上了解机体的抗脂质过氧化作用。     

NAD-Sepharose 6MB亲和胶的制备及在2,5-二酮基-D-葡萄糖酸还原酶纯化中的应用

目的制备NAD亲和胶,用于分离纯化2,5-二酮基-D-葡萄糖酸还原酶。方法将NAD通过丁二酸二酰肼与溴化氰活化的Sepharose6MB胶联结,制得NAD亲和胶。从棒杆菌细胞中提取的2,5-二酮基-D-葡萄糖酸还原酶粗酶液用NAD亲和胶进行亲和层析纯化。结果NAD的结合率为71．67％;亲和层析回收率为77．27％;酶的比活提高了1．1倍。结论以NAD为配基的亲和胶用于纯化以NADPH为辅酶的2,5-DKG还原酶行之有效。     

双氯酚酸钾片人体生物等效性研究

目的评价双氯酚酸钾实验制剂和参比制剂的生物等效性。方法8名健康男性志愿者交叉单剂量口服双氯酚酸钾实验制剂或参比制剂50mg,采用反相高效液相色谱法测定经时过程血药浓度,血药浓度时间数据用3p97药代动力学实用程序拟合,计算其药代动力学参数。结果实验制剂和参比制剂主要药代动力学参数Ka分别为(3．042±1．356)h     

煤快速加氢热解焦油组成的分析

采用溶剂萃取－化学处理－柱层析相结合的预处理分离程序、色－质联用和色－红联用结合色谱保留值的定性方法,分析研究了内蒙东胜煤快速加氢热解焦油的化学成分和结构,推测鉴定出２００多种化合物,并对具有代表性的１９种多环芳烃进行了定量分析,讨论了快速加氢热解焦油化学成分的结构特征。     

柱切换HPLC法测定腹水中左旋氧氟沙星的含量

目的：应用柱切换ＨＰＬＣ技术直接测定腹水中左旋氧氟沙得含量。方法：采用磷酸二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁溴化化铵（１００：４）为预处理流动相,经μＢｏｎｄａｐａｋ Ｃ１８柱预处理后,切换到Ｉｒｒｅｇｕｌａｒ＿ＨＣ１８分析柱,以甲醇－磷权二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁基溴化铵（３０：７０：４）为分析流动相进行分离测定,紫外线皮长为２９４ｎｍ。结果：左旋氧氟     

双氯芬酸钠微乳的家兔体内药代动力学考察

目的 :考察双氯芬酸钠微乳在家兔体内的药代动力学过程。方法 :家兔单次口服双氯芬酸钠微乳和双氯芬酸钠混悬液后 ,用 HPL C法测定血中双氯芬酸钠浓度。 结果 :双氯芬酸钠微乳和混悬液的 AU C0 -∞ 分别为 13.45 6和 10 .5 84μg· h· m l- 1 ,cmax 1 为 2 .85 2和 3.145 μg/m l,tmax1 为 1.438和 0 .75 0 h。 结论 :双氯芬酸钠微乳在家兔体内吸收过程较为平缓 ,可在较长时间内维持一定的血药浓度。     

Single-cell fate mapping reveals widespread clonal ignorance of low-affinity T cells exposed to systemic infection

T cell ignorance is a specific form of immunological tolerance. It describes the maintenance of naivety in antigen-specific T cells in vivo despite the presence of their target antigen. It is thought to mainly play a role during the steady state, when self-antigens are presented in absence of costimulatory signals and at low density or to T cells of low affinity. In how far antigen-specific T cells can also remain clonally ignorant to foreign antigens, presented in the inflammatory context of systemic infection, remains unclear. Using single-cell in vivo fate mapping and high throughput flow cytometric enrichment, we find that high-affinity antigen-specific CD8⁺ T cells are efficiently recruited upon systemic infection. In contrast, most low-affinity antigen-specific T cells ignore the priming antigen and persist in the naïve state while remaining fully responsive to subsequent immunization with a high-affinity ligand. These data establish the widespread clonal ignorance of low-affinity T cells as a major factor shaping the composition of antigen-specific CD8⁺ T cell responses to systemic infection.     

猪血小板衍化生长因子的提取和纯化

目的 建立一种简单、经济的血小板衍化生长因子（PDGF）的纯化方法。方法 血小板衍化生长因子（PDGF）具有耐酸性（pH2.5）,耐热（100℃）以及耐受2%SDS的特性。采用酸醇提取、热处理、离子交换层析、分子筛等蛋白分离和纯化技术从猪血小板中提取和纯化PDGF。结果 猪血小板衍化生长因子（pPDGF）纯化倍数达7582倍,活性回收率为3%。结论 本方法简单、经济,纯化的猪血小板衍化生长因子(pPDGF）具有明显的生物学活性。     

Effect of regular training on the anthropometric parameters and urine steroids in childhood

The changes in the anthropometric data and urine steroid metabolites caused by regular training in children in two age groups (11 and 14 years old) were investigated. The skinfolds of older girls participating in regular athletic, swimming or soccer training were thinner compared with age-matched control groups (P < 0.01) and their body mass and constitution were lower (P < 0.05). In the other groups no significant differences were observable in the anthropometric parameters. The trained children in all groups had significantly higher exercise times on the cycle ergometer (P < 0.01, in young boysP < 0.05). The strength of their hands was lower in three trained groups: in younger boys (P < 0.05), in younger girls (P < 0.01) and in older girls (right handP < 0.01, left handP < 0.05). The urinary excretion of androsterone (P < 0.02), 11-ketopregnanetriol (P < 0.01) and pregnenetriol (P < 0.02) was decreased in the older trained girls; pregnenetriol was increased in younger boys (P < 0.05). Urinary excretion of cortisol metabolites was increased in trained boys [in younger boys: tetrahydrocorticosterone (P < 0.05) and 20-hydroxycortisol (P < 0.05); in older boys allotetrahydrocortisol (P < 0.02), cortisol (P < 0.05) and 20-hydroxycortisol (P < 0.05)]. There were no significant differences in the younger girls. In the trained older girls urinary excretion of cortisol metabolites was decreased: tetrahydrocortisone (P < 0.02), allotetrahydrocorticosterone (P < 0.01), tetrahydrocortisol (P < 0.05), -cortolone (P < 0.01), cortisol (P < 0.02), 6-hydroxycortisol (P < 0.01) and 20-hydroxycortisol (P < 0.05). A multivariate analysis of the data from the trained groups and sedentary, age-matched control groups showed that regular training has a significant effect on steroid excretion.     

Analytical Chiral Separation of the Stereoisomers of a Novel Carbonic Anhydrase Inhibitor and Its Deethylated Metabolite,and the Assignment of Absolute Configuration of the Human Metabolite and Chiral Degradation Products

Several approaches to the separation of four stereoisomers, 1–4, of a novel, topically active, carbonic anhydrase inhibitor, 1, with two chiral centers in the molecule and four isomers, 5–8, of its chiral metabolite, 5, were evaluated. These methods include nonchiral derivatization followed by separation on chiral stationary phases (CSPs) and chiral derivatization and separation on nonchiral columns and on CSPs. Baseline separation of stereoisomers 1–4 was achieved in less than 15 min after chiral derivatization with (S)-(+)-l-(l-naphthyl)ethyl isocyanate (NEIC) and chiral chromatography on a (R)-N-(3,5-dinitrobenzoyl)phenyl glycine (DNBPG) column under normal phase (NP) conditions. Similarly, isomers 5-8 were baseline separated in less than 20 min after derivatization with NEIC and chromatography on nonchiral (nitrophenyl) and chiral [(S)-(3,5-dinitrobenzoyl)leucine; DNBL] columns in series under the same NP chromatographic conditions. Only partial separation of the diastereomeric derivatives was observed on a variety of nonchiral columns. In addition, all other direct and indirect chiral separation approaches gave only partial separation of at least two stereoisomers within the group of 1–4 or 5–8. The details of chiral separations using various methods and separation () and capacity factors (k) of the derivatized isomers 1–8 on a series of chiral and nonchiral columns are presented. Using these methods, the absolute configuration of the human metabolite of 1 was established as S ₁ S ₂ (5), and the heat (HD) and light (LD) degradation products of 1 as R ₁ S ₂ (3) and S₁ S ₂ (5), respectively.