血清miRNA提取方法的优化

利用正交试验法优化血清miRNA的提取方法,为进一步研究血清miRNA作为生物标记物提供方法学基础.本文以hsa-miRNA-223-3p,hsa-miRNA-483-5p和hsa-miRNA-16为检测目标,通过比较Trizol法和Trizol+酚/氯仿法的miRNA提取效率和qPCR检测结果,应用正交试验选择出最优实验方案.结果表明血清miRNA的最优提取方法为:血清用DEPC水4倍稀释,离心条件为4℃,12 000r·min-1,离心时间为10min,Trizol法提取.     

叶绿素衍生物CPD4在人血清中的吸收光谱

研究了不同浓度（１０、５０μｇ／ｍｌ）的叶绿素衍生物４号（ＣＰＤ４）在含１０％不同血清人血清（分Ａ、Ｂ、Ｏ型三种）生理盐水中以及纯蒸馏水中的吸收光谱，并对结果进行了分析和比较，该工作对ＣＰＤ４在ＰＤＴ临床上的应用具有一定的意义。．     

Effect of volatile anesthetics on the circular dichroism of bilirubin bound to human serum albumin

The characteristic circular dichroism of bilirubin bound to human serum albumin undergoes a remarkable sign inversion on addition of halothane, chloroform and other volatile anesthetics. This sign inversion, which is completely reversed by removal of the anesthetic, reflects a pronounced conformational change of the bound ligand; probably a complete inversion of chirality. The observation suggests that association of volatile anesthetics with proteins can markedly alter the internal topography of receptor sites and potentially influence the stereoselectivity of ligand binding.     

完全体外化牛细胞核移植的研究

本研究探讨了用体外成熟卵母细胞和体外受精胚胎进行核移植的可能性。本研究结果在我国首次获得牛核移植的成功。从屠宰场回收的卵巢中抽取卵球－卵母细胞复合体，采用以前报道的方法（石德顺等，广西农业大学学报’１９９４：１３：１－５）进行体外成熟（ＩＶＭ）和体外受精（ＩＶＦ），获得ＩＶＭ卵母细胞和ＩＶＦ胚胎，体外成熟２３－２４ｈ后，在含０．１％透明质酸酶的无钙镁ＰＢＳ（ＰＢＳ）内，用细管反复吹打去掉卵丘细胞。选择具有明显第一极体的卵母细胞通过显微操作移去第一极体及其附近约一半的细胞质进行去核，去核的卵细胞在成熟液内继续培养到ＩＶＭ３０ｈ．体外发育到８－３２细胞期的ＩＶＦ胚胎用作核供体．供体胚胎用含０．２５％Ｐｒｏｎａｓｅ（溶于ＰＢＳ￣－）溶解透明带，并用细管吹打将其分散成单个卵裂球，在ＩＶＭ３０ｈ将单个卵裂球显微注入去核卵母细胞的卵周隙内，并在ＩＶＭ３１ｈ用８０Ｖ／ｍｍ４０μｓ两次电脉冲（间隔１秒）来诱导卵裂球与去核卵母细胞的融合。融合卵在颗粒细胞单层培养滴内共培养，体外培养２４ｈ后观察卵裂结果，经体外培养５－８天后发育到桑椹和囊胚的核移植胚胎移植到同期发情的受体．２个月后直检妊娠情况．本实验中共操作１９４枚卵母细胞，     

Preferential excretion of glycated albumin in C57BL-Ks-J mice: Effects of diabetes

Urinary excretion of glycated albumin was quantitated in genetically hyperglycemic mice (C57BL-Ks-J, db/db mice), a model for non-insulin-dependent diabetes mellitus, and compared with their non-diabetic littermates. The data indicated a preferential excretion of glycated albumin in non-diabetic mice. This phenomenon of editing of glycated albumin is decreased significantly in diabetic mice. Quantitative measurements of overall excretion of glycated albumin suggested that the loss of editing in diabetic mice is due to the dilution of glycated albumin by the unmodified albumin which is excreted in large amounts in diabetic mice. Therefore, the loss of editing observed in this model resembled the one we characterized in insulin-dependent diabetic humans and a streptozotocin-diabetic rat model³.     

Purification of platelet-derived endothelial cell growth inhibitor and its characterization as transforming growth factor-β type 1

In 1986, Brown and Clemmons (Proc. natl Acad. Sci. USA83 (1986) 3321) showed that platelets contain a substance, platelet-derived growth inhibitor (PDGI), that inhibits in vitro endothelial cell replication. Although platelets are rich in transforming grwoth factor (TGF-), PDGI was considered not to be related to TGF-, on the basis of its reported properties (extraction from platelets at neutral pH, binding to heparin-Sepharose). However, we purified PDGI to near homogeneity and showed that on the basis of HPLC retention behavior, in vitro growth inhibitory activities with several cell types, receptor binding, and immunoneutralization of growth inhibitory activity with specific anti-TGF- type 1 antibodies, PDGI is most probably identical with TGF- type 1.     

Evaluation of the hepatotoxicological effects of a drug in an in vivo/in vitro model

Both in vivo and in vitro models have certain disadvantages for the study of the chronic hepatotoxicity of drugs. The aim of this work was to evaluate a new approach based on an in vivo/in vitro model. After chronic in vivo treatment of rats with Vincamine and Vindeburnol (an eburnamenine derivative which exhibits hepatotoxic properties in man) liver cells were isolated, and functional and metabolic disorders (metabolic utilization of fructose and protein biosynthesis) were studied to determine injury. The results showed no modification of blood parameters, but a direct relationship between the dose of Vindeburnol administered in vivo and the metabolic disorders observed in vitro, evidencing the high sensitivity and reliability of this model.     

In vitro amiodarone protein binding and its interaction with warfarin

Summary The binding of amiodarone to human plasma protein and to bovine serum, albumin was studied by three different methods, ultracentrifugation, equilibrium dialysis and fluorescence spectroscopy. The fraction of amiodarone bound to plasma protein amounted to 96.3%. The changes in the binding properties of 1-anilino-naphthalene-8-sulfonate for bovine serum albumin using warfarin and amiodarone as independent inhibitors were analyzed in terms of binding site specificity. The findings indicated that amiodarone and warfarin have two different binding sites on bovine serum albumin, so a noncompetitive inhibition mechanism was indicated. On the basis of our data we cannot exclude other mechanisms of interaction besides direct displacement of one drug by another; nevertheless, metabolite interference between amiodarone and coagulation cofactors may better explain the enhancement of warfarin's pharmacological action in association with amiodarone.This work was partially funded by the CNR (National Research Council, Rome, Italy), Program on Clinical Pharmacology and Rare Diseases. The authors would like to thanks Drs E. Marzi and E. riva for their help.     

Determination of certain physical and chemical characteristics of the activating serum factor from preparturient women and women with habitual abortions

Summary Follow-up investigation of the blood sera from preparturient women and women with habitual, abortions showed the presence of a factor which has an activating effect on smooth muscle preparations because it causes the release of prostaglandins. Gel-chromatographic counter flow separation and microelectrophoresis of the blood sera have shown that the isolated serum factor is a water soluble glycopeptide with a molecular weight of about 2000.     

血清金属元素含量与缺血性脑血管病相互关系探讨

对缺血性脑血管病（ＩＣＶＤ）患者血清Ｚｎ、Ｃｕ、Ｃｒ、Ｍｇ元素进行测定,结果表明患者血清Ｃｕ、Ｃｒ、Ｍｇ含量比对照组低,Ｚｎ含量和Ｚｎ／Ｃｕ比值比对照组高,测定患者Ｚｎ／Ｃｕ比值较测定单一元素Ｚｎ、Ｃｕ更有意义。