大鼠肾小球微循环活体观察实验研究

结扎大鼠一侧输尿管,复制肾盂积水模型。在冷光源－光导纤维－导光棒的透射照明下,通过显微电视系统,成功地观察到完整肾脏皮质表层肾小球及其周围血管微血流动态的清晰图像。正常肾小球呈树状分布,密集、大小均匀。高倍镜下可清楚辨认入球及出球小动脉,其血细胞流态为直线状。并发现肾小球有交替开放现象。     

血循环障碍在高原冻伤中的形态计量观测

本文对高原冻伤中血液循环障碍作形态计量,旨在探讨血循环障碍在冻伤过程中的变化及高原冻伤发病机理中所起的作用。实验选用Ｗｉｓｔａｒ雄性大鼠４０只,随机分为平原冻伤组、急性低氧冻伤组和低氧习服冻伤组。习服组动物于低压舱内模拟海拔６０００ｍ缺氧每日４ｈ,连续两周。其余动物常规饲养。习服期满次日习服组与低氧组一同进入舱内模拟海拔６０００ｍ低氧４ｈ,再行冷冻。冻后继续低氧４ｈ。冻后４８ｈ取材。对各组动物冻后４８ｈ冻肢皮下血管的病变作图象分析。结果发现,平原组血管淤滞、血栓绝对数及其百分比均为最低,习服组最高,低氧组居中。但低氧组与平原组的血栓／淤滞百分比无明显差别。骨骼肌坏死的面积百分比习服组显著高于低氧组与平原组,而后两组间无差别。血栓／淤滞百分比与骨骼肌坏死面积百分比之间的有高度相关关系。冻融是直接引起血管内皮损伤的原发因素,局部血液循环障碍是造成严重的继发损伤的主要原因。     

Acyl Group Migrations in 2-Monoolein

Acyl migration in 2-monoolein dissolved in solvents under conditions common in lipid modification reactions has been studied. The effects on acyl migration of solvent, incubation temperature, water activity, polar additives and solid additives have been investigated. Extensive acyl migration occured in aliphatic hydrocarbons and water-miscible alcohols under dry conditions. The acyl migration rate could be decreased in several nonpolar solvents by adding a small amount of water or an alcohol. Increasing water activity had no effect in isooctane, but decreased the acyl migration rate dramatically in methyl tert-butyl ether and methyl isobutyl ketone. Several commonly used enzyme supports catalysed acyl migration, showing that supports with surface charges could catalyse acyl migration.     

PLD activation in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA

Pertussis toxin-insensitive GTP-binding protein was observed to be involved in prostaglandin F2α(PGF2α)-induced phosphoinositide metabolism in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA (CHO-PGF2α·R cells) (Ito, S. et al. Biochem. Biophys. Res. Commun. 200: 756, 1994). In the present study, we investigated PGF2α-induced PLD activation in CHO-PGF2α·R cells. PLD activation was examined by measuring the production of [³H]phosphatidylbutanol ([³H]PBut), a specific product of the PLD-catalyzed transphosphatidylation reaction. PGF2α-induced [³H]PBut formation was concentration-dependent with the maximal level obtained at 1 μM PGF2α. The maximal [³H]PBut formation was observed at 2 min after addition of PGF2α. Depletion of extracellular Ca²⁺ with EGTA suppressed PGF2α-induced PLD activation by 50%. PKC inhibitors Ro31–8425 and calphostin C inhibited PGF2α-induced [³H]PBut formation by 50%. PTK inhibitors genistein and herbimycin A failed to inhibit PGF2α-induced PLD activation. A combination of maximal effective concentrations of PGF2α (1 μM) and PMA (100 nM) enhanced PLD activation in an additive manner. Pretreatment of the cells with PMA for 2 h down-regulated PKCα and decreased PGF2α-induced PLD activation. These results suggest that PLD activation by PGF2α is mediated by both PKC-dependent and -independent pathways and that PKCα is involved in the former pathway.     

Characterization of rapeseed myrosinase-binding protein

Myrosinase-binding proteins (MBPs) were purified from seeds of Brassica napus L. (oilseed rape). The proteins were characterized with respect to amino-acid composition, peptide sequence and isoelectric points. Gel electrophoresis and Western blotting of protein extracts from mature seeds showed the existence of at least ten proteins reacting with a monoclonal anti-MBP antibody and ranging in molecular size from 110 to 30 kDa. Proteins other than MBP reacting with the anti-MBP antibody were assigned as myrosinase-binding protein-related proteins (MBPRPs). Two MBPRPs were purified by immunoaffinity chromatography and characterized with respect to partial amino-acid sequence. Sequence identities were found between MBP and MBPRP. Western blot analysis of protein extracts from different tissues of B. napus showed that MBPRP is present in the whole plant, whereas MBP mostly occurs in the mature seed. A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to investigate the occurrence of MBP and MBPRP in developing seeds of some species in the Brassicaceae family.Abbreviations FPLC fast protein liquid chromatography - MBP myrosinase-binding protein - MBPRP myrosinase-bindingprotein-telated protein - PBS phosphate-buffered saline     

Temperature and dissolved oxygen concentration as parameters of Azotobacter chroococcum cultivation for use in biofertilizers

Summary Azotobacter chroococcum was grown in continuous culture at two temperatures (30 °C and 20 °C) and different dissolved oxygen tensions (DOT) (30 % to 40 % and 70 % to 80 % of air saturation), respectively. At the temperature of 30 °C and low DOT a relatively high volumetric productivity and efficiency of nitrogen fixation were obtained. After lowering the temperature to 20 °C, an intensive formation of cysts was observed associated with a drastic decrease of the bacterial growth. Bacteria in the form of cysts kept their physiological activity for a long period of time depending on temperature and preparation.     

创伤后巨噬细胞对T细胞白介素2及白介素2受体α基因表达的直接接触抑制作用

以25μg/ml的丝裂霉素C处理巨噬细胞30min,可阻断巨噬细胞白介素1(IL-1)、白介素6(IL-6)、肿瘤坏死因子(TNF)及前列腺素E_2(PGE_2)的合成与分泌。创伤小鼠巨噬细胞经丝裂霉素C处理后,可明显抑制正常T细胞白介素2(IL-2)mRNA及IL-2受体(IL-2R)αmRNA水平,并增强Ts细胞的抑制活性。去除T细胞中Ts细胞可使巨噬细胞的抑制作用消失。表明创伤后巨噬细胞可通过直接的细胞接触方式抑制T细胞IL-2及IL-2 Rα的基因表达,且这一作用是通过增加Ts细胞活性而实现的。     

食管癌病人运铁蛋白(Tf)和组特异性成份(Gc)亚型分布的研究

应用超薄层聚丙烯酰胺凝胶等电聚焦电泳法, 分析了潮汕地区216例无血缘关系、临床上诊断为食管癌的病人和216例健康人的运铁蛋白(Tf)亚型分布情况, 结果发现:食管癌病人组TfC1C1纯合子频率为0.2639,Tf*Cl基因频率为0.4745,显著低于正常人组(分别为0.4352和0.6227,均为P<0.0 01);同时,食管癌病人组TfC2C2纯合子频率为0.2278,Tf*C2基因频率为0. 4977,显著高于正常对照组(分别为0.1852,P<0.05,和0.3634,P<0.001)。应用超薄层聚丙烯酰胺凝胶等电聚焦结合免疫固定法,分析了潮汕地区21 7例无血缘关系的临床上诊断为食管癌病人和 217例健康人的组特异性成份(Gc)亚型的分布,发现两组间无显著性差异。     

Structural and immunochemical identification of Leb glycolipids in the plasma of a group O Le(a-b-) secretor

Total non-acid glycosphingolipids were isolated from the plasma of a healthy red blood cell group O Le(a-b-) salivary ABH secretor individual. Glycolipids were fractionated by HPLC and combined into eight fractions based on chromatographic and immunoreactive properties. These glycolipid fractions were analysed by thin-layer chromatography and tested for Lewis activity with antibodies reactive to the type 1 precursor (Le^c), H type 1 (Le^d), Le^a and Le^b epitopes. Fractions were structurally characterized by mass spectrometry (EI-MS and LSIMS) and proton NMR spectroscopy. Expected blood group glycolipids, such as H type 1, (Fuc1-2Gal1-3GlcNAc1-3Gal1-4Glc1-1Cer) were immunochemically and structurally identified. Inconsistent with the red cell phenotype and for the first time, small quantities of Le^b blood group glycolipids (Fuc1-2Gal1-3(Fuc1-4)GlcNAc1-3Gal1-4Glc1-1Cer) were immunochemically and structurally identified in the plasma of a Lewis-negative individual. These findings confirm recent immunological evidence suggesting the production of small amounts of Lewis antigens by Lewis negative individuals. Abbreviations: HPLC, high performance liquid chromatography; TLC, (high performance) thin layer chromatography; EI-MS, electron impact ionisation mass spectrometry; LSIMS, liquid secondary ion mass spectrometry; NMR, nuclear magnetic resonance spectroscopy. The sugar types are abbreviated to Hex for hexose, HexNAc forN-acetylhexosamine and dHex for deoxyhexose (fucose). The ceramide types are abbreviated to d for dihydroxy and t for trihydroxy base, n for non-hydroxy and h for hydroxy fatty acids; LCB, long chain base.     

腺苷酸激酶基因在大肠杆菌中的可溶性高表达

报道了鸡肌腺苷酸激酶基因的克隆和在温控启动子P_RP_L调控下在大肠杆菌中的可溶性高效表达.SDS-PAGE分析表明,鸡肌腺苷酸激酶的含量可占大肠杆菌细胞总蛋白含量的38％.利用Johnson等的干冰/乙醇-冰水浴反复冻融法,可将此重组蛋白进行富集,纯度可达85％以上.鸡肌腺苷酸激酶可与抗兔肌腺苷酸激酶单克隆抗体产生强的交叉反应.