小鼠心肌组织3种总RNA提取方法比较

目的:探索一种高效的心肌组织总RNA提取方法,对以后总RNA提取工作进行指导。方法:取小鼠心肌组织,分别采用剪刀剪碎法(剪碎法)、研钵加液氮研磨法(研磨法)、离心柱法(过柱法)对小鼠心肌组织进行总RNA提取,紫外分光光度计定量,聚丙烯酰胺凝胶电泳初步定性。结果与结论:研磨法提取出的总RNA量大,质量可靠,较适合于日常总RNA提取;剪碎法对于一些实验条件较差的地区仍然是一种可取的方法;过柱法对于小量心肌组织的提取较为适合。3种方法均能提取出总RNA。     

克山病区服锌人群高密度脂蛋白胆固醇含量探讨

目的 将单一显色法代替酶法测定克山病病区服锌人群血清高密度脂蛋白胆固醇含量。方法 应用沉淀剂将血清中的低密度和极低密度脂蛋白胆固醇沉淀后，采用酶法和单一显色法对比测定上清液中胆固醇含量。结果 单一显色法精密度试验，变异系数为4.5%、回收试验其平均回收率为98.6%；30份血清样品使用酶法和单一显色法对比测定差异无显性（P＞0.5)。结论 单一显色法完全可代替酶法，为一种简易、准确的测定血清高密度脂蛋白胆固醇含量的方法。     

高甘油三脂血症的研究进展（续二）

六、实验室检查 (一)血浆(血清)甘油三脂测定 1.湿化学法:酶法是临床常规的首选法,非酶法已不再使用。酶法:①比色法;②紫外线(UV)分光光度法;③不受游离甘油影响的结合法。前二种是基于脂酶的一步反应,即水解TG定量产生游离甘油;后一种是在一步反应中破坏原来存在的游离甘油,然后第二步才是酯酶反应。如用前二种方法所测得TG总值中减去0 11mmol/L(10mg/dl)或总TG值的7%作为粗略的平均误差。如某些情况(糖尿病,肾     

多指标综合评价艾滋病口腔含漱液的提取工艺

目的：优选爱口含漱液的提取工艺,保证制剂质量。方法以三因素（加水量、提取时间和提取次数）为考察因素,按三个指标（盐酸小檗碱含量、总多糖含量和浸膏率）采用综合评价方法考查爱口含漱液的最佳提取工艺。结果该制剂最佳提取工艺为加水量6倍,提取时间30 min,提取3次。结论优选最佳提取工艺方案可行,稳定性好,能有效保证制剂质量。     

化瘀无糖颗粒制备工艺的筛选

目的优选化瘀无糖颗粒的制备工艺,研制无糖剂型。方法采用正交试验法,以干膏量和咖啡酸含量的综合评价为指标进行提取工艺筛选;以颗粒成型性、吸湿性、溶化性为指标确定糊精用量。结果确定的煎煮条件为加10倍量水煎煮3次,2 h/次;糊精用量为干浸膏量的10%;阿斯巴甜用量为粉末总量的0.3%。结论采用加10倍量水煎煮3次、2 h/次,加入干浸膏量10%的糊精、粉末总量0.3%的阿斯巴甜,干法制粒工艺能成功制备化瘀无糖颗粒,且该制备工艺稳定可行,所制得化瘀无糖颗粒各质量指标均符合《中国药典》2010年版规定。     

盐酸小檗碱对AchE抑制作用的实验研究

目的:体外观察盐酸小檗碱对大鼠脑组织和血清中乙酰胆碱酯酶(AchE)活性的抑制作用.方法:采用现代分子模拟技术,体外模拟以盐酸小檗碱为代表的5种生物碱(配体)与AchE(受体)相互分于对接效应与差异性;大鼠腹主动脉取血后断头处死,制备脑组织匀浆,采用双缩脲法测定脑组织匀浆中总蛋白含量,用酶法检测AchE活性,采用生物学...     

苦参总碱对血管平滑肌细胞增殖及细胞周期的影响

目的 探讨苦参总碱对血管平滑肌细胞(VSMC)增殖及细胞周期的影响.方法 MTT法检测不同浓度苦参总碱对VSMC增殖的影响;考马斯亮蓝法测定细胞总蛋白含量;采用流式细胞仪分析细胞周期.结果 与对照组相比,苦参总碱对VSMC增殖具有抑制作用,该作用呈浓度依赖性;流式细胞术检测结果表明,苦参总碱可以使处于Go/G1期的VSMC百分比增高.结论 苦参总碱是一种有效的抗VSMC增殖的药物.     

类风湿性关节炎细胞水平建模四种方法比较

目的 比较四种方法 在培养类风湿性关节炎(RA)滑膜细胞(FLS)中的异同,为研究RA提供较好的体外实验模型建模方法 .方法 20例滑膜组织标本均用组织块法、单胶原酶消化法、单胰酶温消化法和双酶消化法四种方法 分离、培养RA FLS,均取培养第7日细胞计数,取第4代细胞鉴定,免疫组化鉴定细胞,测定分离细胞的数量和纯度.结果 四种培养方法 的培养时间比较,单胶原酶法>双酶消化法>组织块法>单胰酶法(P＜0.05).组织块法及双酶消化法所得细胞数较单胶原酶法及双酶消化法少(P＜0.05).第4代细胞均以FLS为主(>95%).结论 四种方法 体外培养可获得生物学特性稳定的RA FLS细胞系,其中组织块法是细胞水平建立RA体外实验模型成功率较高的方法 ,单纯胰酶法是较快速的方法 .     

痰结核分支杆菌培养前处理碱-酶法的探讨

本文采用多酶液+2％NaOH(碱-酶法)作消化液,对288份痰标本进行前处理,接种于酸性罗氏培养基。结果:阳性率(45.8％)较对照组2％NaOH前处理法(碱法)的阳性率(35.1％)高、污染率低、菌株生长时间快。     

延胡索总碱提取方法研究

目的 :改进延胡索总碱的提取方法。方法 :采用正交法 ,考察提取方法、提取溶媒、酸碱度等因素 ,以延胡索乙素为考察指标 ,用 HPL C法测定延胡索乙素的含量 ,比较延胡索总碱提取工艺。结果 :酸碱度对延胡索乙素的得率有极显著影响 ,回流提取好于渗漉 ,醇提好于水提。结论 :采用碱酸度为 PH4.5的 HAC乙酸溶液回流提取最佳     

广西广州管圆线虫病疫源地调查

目的了解广西广州管圆线虫病疫源地的分布现状,为预防控制广州管圆线虫病提供科学依据。方法采用区域抽样法,由中国疾病预防控制中心统一抽取调查点。从每个点的野外、螺类养殖场、餐饮场所、自由市场等收集各类软体动物,采用酶消化法检查,阳性者计幼虫数。结果采集到的软体动物有福寿螺、褐云玛瑙螺、蜗牛和蛞蝓等4类。每个调查点都发现有阳性中间宿主,蛞蝓、福寿螺、玛瑙螺和蜗牛平均感染率分别为11.71%,10.82%,8.85%和2.67%,阳性者平均感染度分别为231条,224条,38条和2.5条幼虫。结论广西软体动物种类多、分布广,广州管圆线虫病疫源地广泛存在,中间宿主感染率和感染度高,发生疫情的可能性大,应该采取相应的防控工作。     

脂质体包被冬虫夏草多糖体对慢性活动性乙型肝炎的疗效观察

对55例乙型慢性活动性肝炎患者,30例采用脂质体包被的冬虫夏草多糖体冶疗,25例用单纯冬虫夏草多糖体治疗。结果肝功能在两组都有明显的改善,调整了免疫CD4/CD8及NK。在治疗组由0.97与13.4％提高到1.23与19.5％,在对照组由0.98与13.68％升到1.11及16.2％,HBeAg及HBV—DNA的消失率在治疗组分别为53.2％及68.4％,优于对照组的24％及30％。一年后对治疗组进行随访,HBeAg与HBV—DNA消失率分别又提高到66.6％及73.6％。提示脂质体包被的冬虫夏草多糖体优于单纯的冬虫夏草多糖体,而且在调整免疫后,HBeAg及HBV—DNA的消率失均有持续的提高。     

Pomegranate juice sugar fraction reduces macrophage oxidative state, whereas white grape juice sugar fraction increases it

The antiatherogenic properties of pomegranate juice (PJ) were attributed to its antioxidant potency and to its capacity to decrease macrophage oxidative stress, the hallmark of early atherogeneis. PJ polyphenols and sugar-containing polyphenolic anthocyanins were shown to confer PJ its antioxidant capacity. In the present study, we questioned whether PJ simple or complex sugars contribute to the antioxidative properties of PJ in comparison to white grape juice (WGJ) sugars. Whole PJ decreased cellular peroxide levels in J774A.1 macrophage cell-line by 23% more than PJ polyphenol fraction alone. Thus, we next determined the contribution of the PJ sugar fraction to the decrease in macrophage oxidative state. Increasing concentrations of the PJ sugar fraction resulted in a dose-dependent decrement in macrophage peroxide levels, up to 72%, compared to control cells. On the contrary, incubation of the cells with WGJ sugar fraction at the same concentrations resulted in a dose-dependent increment in peroxide levels by up to 37%. The two sugar fractions from PJ and from WGJ showed opposite effects (antioxidant for PJ and pro-oxidant for WGJ) also in mouse peritoneal macrophages (MPM) from control as well as from streptozotocin-induced diabetic Balb/C mice. PJ sugar consumption by diabetic mice for 10 days resulted in a small but significant decrement in their peritoneal macrophage total peroxide levels and an increment in cellular glutathione content, compared to MPM harvested from control diabetic mice administrated with water. In contrast, WGJ sugar consumption by diabetic mice resulted in a 22% increment in macrophage total peroxide levels and a 45% decrement in cellular glutathione content. Paraoxonase 2 activity in macrophages increases under oxidative stress conditions. Indeed, macrophage paraoxonase 2 activity was decreased after PJ sugars supplementation, but increased after WGJ sugars supplementation. We conclude that PJ sugar fraction, unlike WGJ sugar fraction, decreases macrophage oxidative state under normal and under diabetic conditions. These antioxidant/antiatherogenic effects could be due to the presence of unique complex sugars and/or phenolic sugars in PJ.     

湖北钉螺基因组5种抽提方法的比较

目的 5种方法提取湖北钉螺基因组DNA,PCR扩增线粒体基因组的COⅡ基因以比较其扩增效果,从而选择适合湖北钉螺大样本基因组抽提的最佳方法。方法将50只钉螺分为5组,每组10只,分别用OMEGA基因组DNA抽提试剂盒、天根海洋动物组织基因组DNA提取试剂盒、简化基因组抽提方法、CTAB法和饱和酚抽提法抽提单只湖北钉螺基因组DNA,从消化、抽提时间、抽提浓度、成本等方面评价各种方法的优劣;PCR扩增线粒体基因组COⅡ基因,测定目的条带相对浓度以衡量扩增效果。结果 5种方法均能提取基因组DNA,消化时间最短的为饱和酚法,其余均为3 h;抽提时间最短的为两种试剂盒和CTAB,最长的为简化法;抽提成本最高的为OMEGA试剂盒法,最低的为CTAB法。5种方法PCR均扩增出COⅡ基因,但目的条带浓度有一定差异,其中OMEGA公司和天根公司试剂盒及实验室简化法提纯的模板PCR扩增浓度较高。结论用OMEGA公司生产的基因组DNA抽提试剂盒抽提湖北钉螺基因组DNA效果好,但成本较高,适用于对小样本的抽提;简化法从提纯效果、成本等方面评价,较适用于大样本抽提。     

A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

AIM： To develop a simple and convenient method for extracting genomic DNA from intestinal microflora for enterobacterial repetitive intergenic consensus （ERIC）-PCR detection.
METHODS： Five methods of extracting bacterial DNA, including Tris-EDTA buffer, chelex-100, ultrapure water, 2% sodium dodecyl sulfate and 10% Triton-100 with and without sonication, were compared with the commercial fecal DNA extraction kit method, which is considered as the gold standard for DNA extraction. The comparison was based on the yield and purity of DNA and the indexes of the structure and property of micro-organisms that were reflected by ERIC-PCR.
RESULTS： The yield and purity of DNA obtained by the chelex method was similar to that obtained with the fecal DNA kit. The ERIC-PCR results obtained for the DNA extracted by the chelex method and those obtained for DNA extracted with the fecal DNA kit were basically the same.
CONCLUSION： The chelex method is recommended for ERIC-PCR experiments in view of its simplicity and costeffectiveness; and it is suitable for extracting total DNA from intestinal micro-organisms, particularly for handling a large number of samples.     

Lignin content in natural Populus variants affects sugar release

The primary obstacle to producing renewable fuels from lignocellulosic biomass is a plant's recalcitrance to releasing sugars bound in the cell wall. From a sample set of wood cores representing 1,100 individual undomesticated Populus trichocarpa trees, 47 extreme phenotypes were selected across measured lignin content and ratio of syringyl and guaiacyl units (S/G ratio). This subset was tested for total sugar release through enzymatic hydrolysis alone as well as through combined hot-water pretreatment and enzymatic hydrolysis using a high-throughput screening method. The total amount of glucan and xylan released varied widely among samples, with total sugar yields of up to 92% of the theoretical maximum. A strong negative correlation between sugar release and lignin content was only found for pretreated samples with an S/G ratio < 2.0. For higher S/G ratios, sugar release was generally higher, and the negative influence of lignin was less pronounced. When examined separately, only glucose release was correlated with lignin content and S/G ratio in this manner, whereas xylose release depended on the S/G ratio alone. For enzymatic hydrolysis without pretreatment, sugar release increased significantly with decreasing lignin content below 20%, irrespective of the S/G ratio. Furthermore, certain samples featuring average lignin content and S/G ratios exhibited exceptional sugar release. These facts suggest that factors beyond lignin and S/G ratio influence recalcitrance to sugar release and point to a critical need for deeper understanding of cell-wall structure before plants can be rationally engineered for reduced recalcitrance and efficient biofuels production.     

日本血吸虫成虫核糖核酸提取方法的研究

本文应用酸性异硫氰酸胍-酚-氯仿一步法从日本血吸虫成虫提取核糖核酸(RNA),并与胍盐/热酚法、胍盐/氯化铯超速离心法的提取结果进行比较。3种方法所分离的RNA,在纯度及完整性上均较满意,但其得率以异硫氰酸胍-酚-氯仿一步法为高,此法简单易行,既经济又省时,对来源不易的少量组织,尤为相宜。     

Effects of diets high in simple sugars on urinary chromium losses

Thirty-seven subjects, 19 men and 18 women, consumed reference diets for 12 weeks formulated by nutritionists to contain optimal levels of protein, fat, carbohydrate, and other nutrients; the following 6 weeks, subjects consumed high sugar diets. The reference diets contained 35% of total calories from complex carbohydrates and 15% from simple sugars while the high sugar diets contained 15% complex carbohydrates and 35% simple sugars. Chromium contents of the reference and high sugar diets were both approximately 16 micrograms per 1000 calories. Compared to the reference diets, consumption of the high sugar diets increased urinary Cr losses from 10% to 300% for 27 of 37 subjects. Urinary Cr excretion of males and females was similar, and there was no significant difference in Cr absorption (calculated from urinary excretion divided by intake times 100) between sexes when adjusted for the increased caloric intake of males. These data demonstrate that consumption of diets high in simple sugars stimulates Cr losses; this coupled with marginal intake of dietary Cr may lead to marginal Cr deficiency, which is associated with impaired glucose and lipid metabolism.     

不同破壁方法提取结核分枝杆菌总RNA的比较

目的 建立简便有效的结核分枝杆菌（Mycobacterium tuberculosis,Mtb）总RNA提取方法。 方法 采用5种方法,分别为160、240和400 W超声波联合Trizol法（简称“160 W超声法”、“240 W超声法”和“400 W超声法”）、玻璃珠（简称“玻珠”）联合Trizol法（简称“玻珠法”）和Trizol法（简称“不加玻珠法”）,对结核分枝杆菌标准株H37Rv进行破壁,提取总RNA。通过荧光定量逆转录PCR评价不同破壁方法对Mtb总RNA提取效果的影响,以灭菌超纯水代替模板为阴性对照。每种方法分别提取3份样本进行检测。最后结果为3份标本的平均值。 结果 玻珠法检测组Ct值为20.67,△Rn值为0.644;400 W超声法检测组Ct值为22.09,△Rn值为0.571;240 W超声法检测组Ct值为21.86,△Rn值为0.503;160 W超声法检测组Ct值为25.21,△Rn值为0.411;不加玻珠法检测组Ct值为28.40,△Rn值为0.299;阴性对照Ct值为40.00,△Rn值为0.000。 结论 直观地从试验结果来看,玻珠法对结核分枝杆菌破壁效果好于超声法。     

Biochemical Analysis of Vagotomized Stomach Wall in Rats

The authors carried out a biochemical analysis of vagotomized rat stomach wall 2–3 weeks after a bilateral vagotomy. The biochemical extraction was performed by the Schneider-Schmidt-Thannhauser method. The phosphate fractions were measured colorimetrically according to Briggs. The nucleic acids were measured on the basis of phosphate and sugar components according to Seibert and Brown. The following compounds were determined: acid-soluble organic and inorganic phosphates, phospholipidphosphate, ribonucleic acidic phosphate, deoxyribonucleic acidic phosphate, phosphoproteid inorganic phosphate, ribonucleic acid and deoxyribonucleic acid from the whole, the glandular and the membranaceous stomach. These were compared with the some constituents of the normal rat stomach. After vagotomy a decrease was observed in acid-soluble inorganic phosphate, in organic acid-soluble phosphate, in phosphoprotein inorganic phosphate of the glandular, membranaceous and the total stomach. The phospholipid phosphate did not decrease. There was no change in the content of ribonucleic acid in the membranaceous stomach, whereas the content of deoxyribonucleic acid showed an increase.