大鼠附睾蛋白质组的提取和二维液相色谱分离

背景与目的：提取犬鼠附睾液蛋白并建立一种利用二维液相色谱法分离附睾蛋白组的方法。方法：分离提取犬鼠附睾液蛋白。样品利用起始缓冲液置换后,进行一维色谱聚焦分离,然后收集pH8．5—4．0之间的组份进行二维反相离压液相色谱分离,最后将获得的二维UV图通过ProteoVue软件转换成PI／UV图谱。结果：成功提取了附睾液蛋白,并通过二维液相色谱成功建立了大鼠头体尾部附睾液蛋白的二维PI／UV图谱,收集了一维色谱聚焦分离的pH8．5—4．0区间的20个组份,并将每个组份进行二维色谱分离后转换为PI/UV图谱。结论：为进一步全面研究附睾蛋白功能和体液差异蛋白质组研究打下了基础。     

曲氏传药中乌头碱的薄层色谱测定

在硅胶Ｇ薄层板上用氯仿：１,２．二氯乙烷：甲醇：氨水（９：１：０．９：０．０７,ｖ／ｖ）为展开剂分离中草药复方曲氏传药中的生物碱,并用改良碘化铀钾试剂显色后对其中的乌头碱进行直接扫描测定。此法简便、快速、准确、灵敏且稳定。     

复方克霉唑乳膏治疗体股癣疗效和安全性评价

复方克霉唑乳膏（compound clotrimazole cream）是一种外用复方制剂,主要成分为1％克霉唑（clotrimazole）、0．05％二丙酸倍他米松（betamethasone dipropionate）和0．1％庆大霉素（gentamicin）。克霉唑为咪唑类广谱抗真菌药物,主要通过作用于真菌细胞膜,使细胞内容物外漏而产生抗真菌作用;二丙酸倍他米松为强效糖皮质激素,具有抗炎、免疫抑制和抗增生作用;庆大霉素为氨基糖甙类抗生素,对革兰阳性菌、革兰阴性菌均有良好的抗菌作用,尤其对金黄色葡萄球菌作用较强。     

叶绿体色素分离实验改进与拓展

针对生物学专业本科生普通生物学实验教学,设计了将薄层色谱法和柱层析色谱法应用于叶绿体色素分离实验.采用硅胶作为吸附材料,以石油醚、苯和丙酮为混合展开剂,用薄层色谱法代替传统的纸色谱法进行叶绿体各色素的分离,用柱层析色谱法进行单体β-胡萝卜素、叶绿素a、叶绿素b和叶黄素制备,实验装置简单,易于操作,特别是各色素条带分离灵敏度高.     

硫酸化分级香菇多糖抗氧化活性研究

提取香菇子实体多糖,用浓硫酸法进行结构修饰,经不同分子量超滤膜超滤分离得到6种不同组份的硫酸化香菇多糖,检测这6种组份抗氧化活性。结果表明,硫酸化香菇多糖具有抗氧化活性,在1-10mg/mL的浓度范围内呈量效关系,且不同分子量组份活性有差异。     

复方壳聚糖消痤乳膏的制备和临床观察

目的制备复方壳聚糖乳膏并用于寻常痤疮的临床观察。方法研制以壳聚糖和苦杏仁酸等为主药的复方消痤乳膏。结果３４例寻常痤疮患者用上述消痤乳膏治疗３周总有效率为９４．１２％。结论消痤乳膏可作为治疗寻常痤疮的一种安全、稳定、有效的制剂。     

复方多糖口服液的制备及质量研究

目的制备复方多糖口服液,并进行定量分析,对各项指标进行检测。方法采用水提醇沉法制备复方多糖口服液,薄层色谱法进行定性鉴别,苯酚-硫酸法测定含量。结果检测波长为490 nm,在4.86～24.7mg范围内多糖呈线性关系,加样回收率为99.84%。结论本法简便易行,结果稳定,可做复方多糖口服液中多糖的含量测定方法。     

嗜麦芽寡养单胞菌临床分布特征与耐药分析

目的了解湖州市中心医院嗜麦芽寡养单胞菌临床分布特征与耐药性。方法采用常规方法分离,用VITE-COMPACT2全自动微生物分析仪进行菌种鉴定,用K—B法进行药敏试验。结果分离到嗜麦芽寡养单胞菌810株,复方新诺明耐药菌株48株（分离率5．9％）。标本来源主要来自ICU室,其次呼吸科,大部分来自痰液标本（约占89．2％）,年龄段以中老年人比率最高。嗜麦芽寡养单胞菌对亚胺培南、美罗培南、头孢吡肟、哌拉西彬他坐巴坦、庆大霉素、妥布霉素、阿米卡星高度耐药;头孢他啶、替卡西林／克拉维酸、环丙沙星耐药率为33．7％～58．2％;头孢哌酮／舒巴坦、左氧氟沙星、米诺环素、复方新诺明耐药率低于30．0％。复方新诺明耐药菌株对头孢哌酮／舒巴坦、左氧氟沙星和米诺环素耐药率分别为60．4％、91．7％和2．0％,对其余抗菌药物耐药率达100．0％。复方新诺明耐药菌株与复方新诺明敏感菌株相比,耐药情况更严重,其中对三、四代头孢菌素、喹诺酮类耐药率显著高于复方新诺明敏感菌株（P〈0．01）;对碳青霉烯类、青霉素类、氨基糖苷类抗菌药物耐药率与复方新诺明敏感菌株相比,差异无统计学意义（P〉0．05）。结论嗜麦芽寡养单胞菌呈高度耐药,对头孢哌酮／舒巴坦、左氧氟沙星、米诺环素、复方新诺明尚敏感,但对复方新诺明耐药的嗜麦芽寡养单胞菌耐药现象更严重。应重视嗜麦芽寡养单胞菌引起的院内感染,尽量减少不必要的侵人性操作,加强抗菌药物的合理规范使用。     

均匀设计法优化绿色木霉菌黄色素薄层色谱展开剂系统

根据系统选择法确定了绿色木霉菌（Trichoderma viride）所产黄色素的薄层色谱展开剂组分,利用均匀设计实验及逐步回归法建立回归模型优化展开剂配比,最终得出优化的展开剂系统为为石油醚-乙酸乙酯-丙酮-乙酸=12．23：1：1．58：0．55,分离效果较理想,所得到的薄层色谱分离函数指标COF2达到了42．09±0．79．     

一种白僵菌代谢产物中生物活性物质的研究——Ⅰ.具有清除自由基？…

通过对白僵菌Ｂｅａｒｖｅｒｉａ ｓｐ．的液体培养及生物活性测定,发现该菌代谢产物个有较强的清除自由基的活性,我们用甲醇成功地提取出该活性成分,同时用色谱等方法对该活性成分进行了分离和制备,并用高压液相色谱法和ＤＰＰＨ薄层试验对其纯讴及活性进行了检验,得到了具消除自由基活性的纯化合物：Ｐ－２４－３。     

Sophoraflavanone G from sophora pachycarpa enhanced the antibacterial activity of gentamycin against Staphylococcus aureus

In this study the enhancement effect of Sophora pachycarpa roots' acetone extract on the antibacterial activity of gentamycin was evaluated against Staphylococcus aureus. Disc diffusion and broth dilution methods were used to determine the antibacterial activity of gentamycin in the absence and presence of plant extract and its various fractions separated by TLC. A clinical isolate of S. aureus was used as test strain. The active component of the plant extract involved in enhancement of gentamycin's activity had Rf = 0.72 on a TLC plate. The spectral data (1H NMR, 13C NMR) of this compound revealed that this compound was 5,7,2',4'-tetrahydroxy-8-lavandulylflavanone (sophoraflavanone G), previously isolated from Sophora exigua. In the presence of 0.03 microg/ mL of sophoraflavanone G the MIC value of gentamycin for S. aureus decreased from 32 to 8 microg/mL (a four-fold decrease). These results signify that the ultra-low concentration of sophoraflavanone G potentiates the antimicrobial action of gentamycin suggesting a possible utilization of this compound in combination therapy against Staphylococcus aureus.     

Radioimmunoassay of estrone sulfate in the serum of normal men after a non-chromatographic procedure that eliminates interference from dehydroepiandrosterone sulfate

Duplicate aliquots of 20 fresh-frozen normal human male sera were prepared for estrone sulfate (ES) radioimmunoassay (RIA) by each of three different methods: the thin-layer chromatography (TLC) procedure we previously reported, a new procedure including overnight heating (100 C) of an ethanol extract reconstituted in dilute acetate buffer, and the new procedure with the hot incubation omitted. The purpose of the 100 C incubation was the selective thermal solvolysis of dehydroepiandrosterone sulfate (DS), the only steroid conjugate present in serum in high enough concentrations to interfere with a high-specificity ES RIA. Dehydroepiandrosterone released by solvolysis and endogenous unconjugated steroids were extracted from the samples with ether before RIA. Estrone sulfate values obtained after the thermal solvolysis preparation averaged 854 +/- 501 pg/ml (SD) versus 826 +/- 474 pg/ml (SD) after the TLC method, with excellent correlation between the two (r = 0.97). Samples prepared by the new method but with thermal solvolysis omitted averaged a 33.8% elevation of measured ES level, an elevation significantly correlated (P less than 0.02) with DS levels obtained from the same specimens. In addition, a single specimen showed no elevation after preparation by the thermal solvolysis method when up to 8 micrograms/ml authentic DS as added before extraction. Compared with the TLC method, the new method also provides substantial savings in specimen volume requirements and sample processing time.     

Quantitative analysis of major dibenzocyclooctane lignans in Schisandrae fructus by online TLC-DART-MS

Introduction – Direct analysis in real time (DART) ion source is a powerful ionising technique for the quick and easy detection of various organic molecules without any sample preparation steps, but the lack of quantitation capacity limits its extensive use in the field of phytochemical analysis. Objective – To improvise a new system which utilize DART‐MS as a hyphenated detector for quantitation. Methodology – A total extract of Schisandra chinensis fruit was analyzed on a TLC plate and three major lignan compounds were quantitated by three different methods of UV densitometry, TLC‐DART‐MS and HPLC‐UV to compare the efficiency of each method. To introduce the TLC plate into the DART ion source at a constant velocity, a syringe pump was employed. The DART‐MS total ion current chromatogram was recorded for the entire TLC plate. The concentration of each lignan compound was calculated from the calibration curve established with standard compound. Results – Gomisin A, gomisin N and schisandrin were well separated on a silica‐coated TLC plate and the specific ion current chromatograms were successfully acquired from the TLC‐DART‐MS system. The TLC‐DART‐MS system for the quantitation of natural products showed better linearity and specificity than TLC densitometry, and consumed less time and solvent than conventional HPLC method. Conclusion – A hyphenated system for the quantitation of phytochemicals from crude herbal drugs was successfully established. This system was shown to have a powerful analytical capacity for the prompt and efficient quantitation of natural products from crude drugs. Copyright © 2010 John Wiley & Sons, Ltd.     

Microbiological study of gentamiycin]

Gentamycin prepared at the All-Union Research Institute of Antibiotics did not differ by its antibacterial spectrum and the activity level from gentamycin samples from other countries. By its activity against clinical strains of Ps. aeruginosa gentamycin was somewhat inferior than polymyxin but much more superior than carbenicillin. An agar-diffusion method using Bac. pumilus NTCC 8241 as the test microbe was developed for determination of gentamycin activity. The gentamycin sulfate complex and the components of gentamycin had the same activity levels, antibacterial spectrum and diffusion capacity.     

Dielectric Barrier Discharge Ionization in Characterization of Organic Compounds Separated on Thin-Layer Chromatography Plates

A new method for on-spot detection and characterization of organic compounds resolved on thin layer chromatography (TLC) plates has been proposed. This method combines TLC with dielectric barrier discharge ionization (DBDI), which produces stable low-temperature plasma. At first, the compounds were separated on TLC plates and then their mass spectra were directly obtained with no additional sample preparation. To obtain good quality spectra the center of a particular TLC spot was heated from the bottom to increase volatility of the compound. MS/MS analyses were also performed to additionally characterize all analytes. The detection limit of proposed method was estimated to be 100 ng/spot of compound.     

Electrochemical interactions between some beta-lactam antibiotics and aminoglycoside antibiotics

Complexation reactions between the two aminoglycosides tobramycin and gentamycin and the two beta-lactam antibiotics carbenicillin and ticarcillin were studied conductimetrically, in aqueous solution. Carbenicillin and gentamycin form a 21 adduct in which about 75% of the antibiotics are bound. Likewise, carbenicillin and tobramycin form a 21 adduct binding about 67% of its components. Tobramycin and ticarcillin also interact, but weakly, binding about 12% of the adduct components. Only a trace of adduct formation was observed between cephalothin and gentamycin and between cephalothin and tobramycin. Cephalothin did not interact with carbenicillin. It appears that the adsorption behavior of the aminoglycosides differs considerably from that of the beta-lactams.     

睫状神经营养因子对听觉损伤的保护作用

本研究以耳廓反射、听觉脑干诱发电位、耳蜗生物电和耳蜗铺片组织学检测为指标,观察重组人睫状神经营养因子对豚鼠庆大霉素耳毒性的防治作用。实验结果表明,睫状神经营养因子能减轻庆大霉素对耳蜗及听神经的损害,具有保护听觉功能的作用。     

两种方案对断奶仔猴慢性肠炎治疗效果的比较

目的探索断奶仔猴慢性肠炎的较优治疗方案。方法比较了治疗方案Ⅰ（庆大霉素碳酸铋联合淮山米粉＋奶粉＋乳酸菌素片）和治疗方案Ⅱ（庆大霉素碳酸铋联合蒙脱石散）对断奶仔猴慢性肠炎的治疗效果。结果治疗方案Ⅰ的治愈率更高,治愈后不易复发,且费用相对低廉。结论此方案可以在生产中推广应用。     

中药制剂研究进展

本文从中药炮制、复方制剂研究和中药质量保证体系三方面评述近几年中药制剂的研究进展。     

Enzymatic synthesis and purification of uridine diphospho-beta-l-arabinopyranose, a substrate for the biosynthesis of plant polysaccharides

Many plant cell wall components such as the polysaccharides xylans and pectins or the glycoproteins arabinogalactan proteins and extensins contain arabinosyl residues. The arabinosyl substituents are thought to be incorporated into these wall polymers by the action of arabinosyltransferases using UDP-l-arabinose as the precursor. UDP-l-arabinose is not commercially available and therefore a procedure for generating UDP-l-arabinose was developed for use in studies on the biosynthesis of the arabinose-containing polymers. In this procedure UDP-d-xylose is incubated with an enzyme preparation from wheat germ and the nucleotide sugars in the reaction mixture are extracted. High-performance anion-exchange chromatography of the extract resolves two major UV-absorbing components: one corresponding to UDP-xylose and a second that elutes earlier. TLC analysis of collected and hydrolyzed fractions demonstrated the presence of l-arabinose in the early eluting fraction. Further analysis by NMR identified the compound as UDP-beta-l-arabinopyranose. The procedure reported here provides an efficient method for preparing either radioactive UDP-l-[(14)C]arabinose or nonradioactive UDP-l-arabinose and can also be used as an assay for UDP-xylose-4-epimerase activity.