大豆制品中胰蛋白酶抑制物活性测定方法的改进——多因素正交试验的应用尝试

本文对胰蛋白酶抑制物活性的测定方法进行了改进。应用多因素正交试验获得了最佳实验条件。最佳实验条件为样品提取液1ml(相当原样品1mg),胰蛋白酶用量4mg,保温时间为37℃15分钟。本文也探讨了胰蛋白酶与光密度的关系,并应用本文的条件和改进的方法对生豆粉和烘烤过的豆粉(150℃30分钟)中胰蛋白酶抑制物的活性进行了测定。     

Diazepam inhibits cholesterol esterification by arterial ACAT and plasma LCAT, in vitro

Benzodiazepine drugs have been reported to have antiatherosclerotic effects in rabbits and roosters and to alter the pattern of circulating lipoproteins in man. The mechanism(s) of these effects has not been elucidated. The studies presented here indicate that diazepam, the most widely used benzodiazepine, is an inhibitor of cholesterol esterification by ACAT in vitro in atheromatous rabbit aortas, in microsomes isolated from atheromatous rabbit aortas, and in normal rat aortas. Diazepam also inhibited LCAT in plasma from man, monkey, rabbit, and rat, in vitro. The ability of diazepam to inhibit these enzyme systems may offer insight into possible in vivo mechanisms of action against atherosclerosis and of lipoprotein modification.     

Dissolving intravenous cyclosporin A in a fat emulsion carrier prevents acute renal side effects in the rat

Cyclosporin A (CyA)-induced nephrotoxicity is still a serious clinical problem. The nephrotoxicity seems to be more pronounced when CyA (solubilized in water with Cremophore EL, as in Sandimmun) is given intravenously than when it is given orally. Using soybean oil in which CyA was dissolved, we prepared an intravenous fat emulsion without an artificial detergent, such as Cremophore EL. The renal effects of our new formula were compared with those of Sandimmun and Cremophore EL in a rat model. CyA in the fat emulsion did not significantly affect the glomerular filtration rate (GFR). Both Sandimmun and Cremophore EL significantly reduced the GFR. These results suggest that a change in the vehicle may obviate the acute nephrotoxic side effects caused by intravenous administration of CyA solubilized by Cremophore EL.     

大豆异黄酮对阿尔茨海默病模型大鼠行为学及抗氧化能力的影响

目的：探讨不同剂量大豆异黄酮（SIF）对β淀粉样蛋白（Aβ）介导的大鼠行为学及抗氧化能力的影响。方法：60只健康雄性SD大鼠随机分为假手术组、模型组、SIF低、中、高剂量组和阳性对照组,每组10只。SIF组按规定剂量灌胃给药,阳性对照组给予维生素E,假手术组和模型组灌胃等量的0.5%羧甲基纤维素钠（CMC-Na）。连续灌胃14d后,模型组和实验组大鼠双侧海马CA1区注射Aβ25-35,假手术组注射等量生理盐水,于术后7d,Morris水迷宫实验检测大鼠学习记忆能力,之后处死大鼠,观察病理形态学改变及检测血清和脑组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧物酶（GSH-Px）、丙二醛（MDA）等抗氧化指标的水平。结果：与模型组比较,SIF各剂量组和阳性对照组平均逃避潜伏期缩短,穿越站台所在位置的次数增多（P〈0.05）,SIF中、高剂量组较阳性对照组潜伏期缩短明显（P〈0.05）。免疫组化结果显示,与模型组比较,SIF各剂量组和阳性对照组可减少海马区Aβ25-35免疫反应阳性产物的表达,SIF高剂量组较阳性对照组减少明显（P〈0.05）。与模型组比较,SIF中、高剂量组及阳性对照组大鼠血清和脑组织SOD和GSH-Px活性不同程度增加,MDA含量不同程度降低（P〈0.05）;SIF高剂量组在提高大鼠血清和脑组织SOD活性及降低MDA含量方面优于阳性对照组（P〈0.05）;SIF高剂量组提高大鼠血清中GSH-Px活性较阳性对照组明显（P〈0.05）,对脑组织中GSH-Px活性的影响与阳性对照组相比无统计学差异（P〉0.05）。结论：SIF能改善Aβ25-35所致阿尔茨海默病模型大鼠的行为能力,可能是通过改善机体氧化还原状态,提高抗氧化水平,由此发挥其神经保护作用。     

Formulation,Characterization, and In Vitro Evaluation of Silymarin-Loaded Lipid Microspheres

The objective of our study was to incorporate and evaluate Silymarin, a chemically defined natural hepatoprotective agent, in lipid microstructured systems. Various constituents of lipid microspheres—namely, internal oily core; surfactant such as soyabean lecithin; and cosurfactants such as span 20, tween 20, tween 80, and propylene glycol—were tried in different concentrations to optimize the final formulation characteristics such as globule size range, structural integrity, sustainability, and percent drug-holding capacity. The final formulation (formulation A) was characterized with respect to size and morphology using transmission electron microscopy and laser diffraction technique. The enhanced mean percent release of 56.70 ± 2.03% was observed in 36 hr from silymarin-loaded lipid microspheres (formulation A), as compared to 18.67 ± 0.192% with silymarin solution (formulation B). Thus, a stable delivery system having synergistic hepatoprotective effect of silymarin and soyabean lecithin could successively be produced for passive targeting to the liver.     

溶血卵磷脂酰基转移酶1自发突变小鼠视网膜组织和闪光视网膜电图改变

目的 观察溶血卵磷脂酰基转移酶1（LPCAT1）自发突变小鼠视网膜组织和闪光视网膜电图（F-ERG）改变情况。方法 Lpcat1基因自发突变纯合子的rd11新生小鼠60只（实验组）和与之同龄的野生型C57BL/6J小鼠60只（对照组）纳入实验。采用免疫荧光法检测两组小鼠视网膜组织中LPCAT1的表达情况。出生后3、6、9 d和2、4、6、8周,行视网膜石蜡切片观察视网膜显微结构;行全视野F-ERG检测,记录暗视杆体反应和明视锥体反应。结果 实验组小鼠视网膜组织中LPCAT1呈阴性表达;对照组小鼠视网膜组织中LPCAT1呈阳性表达,主要分布于视网膜光感受器内节,神经节细胞层可见少量表达。光学显微镜观察发现,实验组小鼠于出生后9 d左右视网膜外核层已基本形成,其光感受器细胞核层数随年龄增长而逐渐减少。对照组小鼠视网膜显微结构正常。F-ERG检测结果 显示,实验组小鼠出生后2、4周暗视杆体系统反应存在,呈降低趋势,出生后6~8周暗视杆体系统反应消失;对照组小鼠各时间点暗视杆体系统反应正常。出生后2周,实验组和对照组小鼠b波振幅分别为（72.8±15.6）、（105.2±21.1） μV,实验组较对照组降低,但差异无统计学意义（t=-2.760,P=0.025）。出生后4周,实验组和对照组小鼠b波振幅分别为（20.6±6.4）、（231.8±32.0） μV,实验组较对照组明显降低,差异有统计学意义（t=-14.471,P=0.000）。实验组小鼠出生后2、4、6周明视锥体系统反应存在,呈降低趋势,出生后8周明视锥体系统反应消失;对照组小鼠各时间点明视锥体系统反应正常。出生后2、4周,实验组小鼠b波振幅分别为（46.8±7.2）、（78.0±8.2） μV;对照组小鼠b波振幅分别为（42.8±6.4）、（91.4±9.4） μV。两组比较,差异均无统计学意义（t=0.930、-2.401,P=0.379、0.043）。出生后6周,实验组和对照组小鼠b波振幅分别为（17.2±2.0）、（116.2±12.9） μV,实验组较对照组明显降低,差异有统计学意义（t=-17.008,P=0.000）。结论 LPCAT1自发突变小鼠视网膜外层光感受器细胞核层数随年龄增长而逐渐减少,F-ERG暗视杆体和明视锥体反应异常。     

Insecticide toxicity to Hyalella curvispina in runoff and stream water within a soybean farm (Buenos Aires, Argentina)

Toxicity to the locally dominant amphipod Hyalella curvispina was assessed in a first-order stream running through a cultivated farm. Cypermethrin, chlorpyrifos, endosulfan and glyphosate were sprayed throughout the studied period. Toxicity was assayed under controlled laboratory conditions with runoff and stream water samples taken from the field under steady state and flood conditions. Ephemeral toxicity pulses were observed as a consequence of farm pesticide applications. After pesticide application, runoff water showed 100% mortality to H. curvispina for 1 month, but no mortality thereafter. Toxicity persistence was shortest in stream water, intermediate in stream sediments and longest in soil samples. Runoff had a more important toxicity effect than the exposure to direct aerial fumigation. The regional environmental features determining fast toxicity dissipation are discussed.     

大豆浓缩磷脂中蛋白质及糖类脱除方法的研究

目的:研究脱除大豆浓缩磷脂中蛋白质及糖类的方法条件。方法:用石油醚和丙酮为溶剂对大豆浓缩磷脂中的蛋白质进行脱除,再用凯式微量定氮法测定蛋白质脱除量。考察磷脂脱蛋白过程中温度、溶剂比、料液比、时间各因素对蛋白脱除量的影响,并通过正交实验对脱除方法进行优化,得到最佳条件。同时用乙醇溶液作为溶剂对大豆浓缩磷脂中的糖类进行脱除,再用蒽酮比色法测定糖的脱除率,考察乙醇浓度、温度对脱除率的影响。结果:在温度为10℃,石油醚与丙酮的体积比为1∶1,料液比为1∶3,静置时间为1h时,蛋白脱除量为302.19 mg蛋白/100g浓缩磷脂;在温度为25℃,乙醇浓度为50%时脱糖率最大,达到81.52%。结论:在上述条件下,脱除蛋白质和糖类后的大豆浓缩磷脂较原料磷脂在透明度上有很大改善。     

钙与大豆异黄酮的联合作用对去卵巢大鼠骨生物力学特性的影响

目的 探讨钙和大豆异黄酮的联合作用对去卵巢大鼠骨生物力学特性的影响.方法 将160只12月龄SD清洁级雌性大鼠随机分为4组,分别为假手术对照(蒸馏水)组、骨质疏松模型(蒸馏水)组、大豆异黄酮(10 mg/ml,染毒容量为3 ml/kg)组、雌激素(0.1 mg/ml,染毒容量为0.1 ml/kg)组,每组再按照加钙(1...     

Novel colloidal associations of soyasaponins and lipid components (DPPC,cholesterol) as potential adjuvants for vaccines

Soyasaponins from soybean (Glycine max) represent promising new potent adjuvants for vaccine research because of their immunostimulating properties and weak hemolytic activity. In the present study, saponin microstructures of soyasaponins (soyasaponin Bb, soyasaponin Ab) with lipid components (cholesterol, DPPC (dipalmitoylphosphatidylcholine)) were designed by the lipid film method. In interaction studies between soyasaponins (soyasaponin Ab/Bb) and Langmuir monolayers (model membranes), composed of cholesterol and DPPC, marked interactions between soyasaponins and a pure cholesterol monolayer were observed. No interaction was detected for soyasaponins with a pure DPPC monolayer. The intercalation of soyasaponins in a mixed DPPC/cholesterol (3:1, w/w) monolayer was only observed for the monodesmosidic soyasaponin Bb whereas the second sugar chain of the bidesmosidic soyasaponin Ab impaired the access to the monolayer. Transmission electron microscopy was used for visualizing particle formation of soyasaponins and lipid components. Pseudo-binary systems (soyasaponin Ab/Bb, cholesterol) formed colloidal associations built up from ring-like subunits in the nanometer size range. In pseudo-ternary systems (soyasaponin, cholesterol, DPPC) soyasaponin Bb attacked the liposomal membrane by forming colloidal associations. Colloidal associations in pseudo-ternary systems with soyasaponin Ab, cholesterol and a phospholipid were only observed in the presence of PE (phosphatidylethanolamine) instead of DPPC. In an MTT assay with a HaCaT cell line (keratinocyte cell line) the cell viability was neither affected by the soyasaponins nor by the corresponding formulations. Both the pure soyasaponin solution and the saponin formulations may be promising adjuvant systems for the intradermal vaccine application. Furthermore, interaction studies between the model antigen ovalbumin and colloidal associations of saponins and cholesterol using MST (Microscale Thermophoresis) gave first indications of an antigen binding to colloidal associations. Ex vivo T-cell proliferation in the presence of soyasaponin Ab was confirmed.