蛹虫草子实体多糖的分离纯化

目的:研究人工培养蛹虫草子实体多糖的分离纯化方法.方法:多糖经脱色、醇析、除蛋白及乙醇分级沉淀,采用Seph-adex G100柱层析纯化.结果:60℃脱色9h脱色率达70.72％.醇析最优条件为:无水乙醇,24h,4倍乙醇.Sevag法抽提30min重复6次除蛋白率达84.09％;抽提lh重复3次、6次除蛋白率分别为81.14％、84.47％,多糖损失率分别为27.03％、43.16％.木瓜蛋白酶除蛋白最优条件为70℃,1:10,2h,除蛋白率为25.67％.粗多糖经乙醇分级沉淀得2种粗多糖成分,分别经Sephadex G100柱层析可进一步纯化为3种较纯多糖成分.结论:采用乙醇分级沉淀及Sephadex G100柱层析纯化粗多糖可得到3种较纯多糖成分,纯化效果较好.     

浒苔多糖的分离、纯化和分析

浒苔（Ｅｎｔｅｒｏｍｏｒｐｈａｐｒｏｌｉｆｅｒａ）经热水提取,Ｓｅｖａｇｅ法除去蛋白质,用乙醇沉淀,ＳｅｐｈａｄｅｘＧ－１００柱层析,得浒苔多糖（简称ＥＰ）精制品。经ＳｅｐｈａｄｅｘＧ－２００柱层析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析未见有核酸和蛋白质的特征吸收峰。总糖含量为８８．８％,其中糖醛酸含量为３３．６％。单糖组成为Ｌ－阿拉伯糖、Ｌ－岩藻糖、Ｄ－甘露糖、Ｄ－半乳糖及Ｄ－葡萄糖,平均分子量为２５０００。     

金针菇深层发酵菌丝体中水溶性多糖的提取条件的研究

本文研究了食用菌深层发酵金针菇菌丝体多糖的提取条件。实验表明,在加水比为１：６,沸水提取４小时,进行三次抽提后,用４倍乙醇沉淀所获粗多糖得率最高。粗多糖经脱色去蛋白后以７０％（Ｖ／Ｖ）乙醇分级可获得最高级分精制多糖。     

响应面法优化北豆根粗多糖的除蛋白工艺及其神经保护活性研究

为研究北豆根粗多糖的最佳除蛋白方法和工艺,探究北豆根粗多糖的神经保护活性。实验比较了酶法、Sevage法、酶-Sevage法、三氯乙酸(TCA)-正丁醇法、酶-TCA-正丁醇法除蛋白效果,筛选出了北豆根粗多糖的最佳除蛋白方法,并用响应面法对该方法进行优化。经响应面分析,以综合评分为指标,考查TCA与正丁醇体积比、北豆根粗多糖溶液与TCA-正丁醇溶液体积比和振摇时间对北豆根粗多糖TCA-正丁醇法除蛋白工艺的影响。用H_2O_2诱导PC12细胞损伤,探究了北豆根粗多糖的神经保护作用。结果表明,北豆根粗多糖的最佳除蛋白方法为TCA-正丁醇法。经响应面优化,确定最优除蛋白工艺为TCA:正丁醇=1∶10.4,北豆根粗多糖溶液:TCA-正丁醇溶液=1∶1.77,振摇时间为36.5 min。采用该工艺对北豆根多糖进行除蛋白,其蛋白清除率为73.1%,综合评分为92.1。神经保护研究结果表明,北豆根粗多糖对H_2O_2诱导的PC12细胞损伤具有明显的保护作用。本研究表明TCA-正丁醇法可以有效的除去北豆根粗多糖中的蛋白质且北豆根粗多糖具有神经保护活性。     

红菇子实体多糖的理化性质及抗癌活性

红菇（Russula vinosa Lindbl）子实体残渣经热水提取,Sevage法除去蛋白,用乙醇沉淀得到粗多糖,经DEAE-Sephadex A-25纯化,得到精制的红菇多糖,经聚丙烯酰胺凝胶（PACE）电泳和高效液相色谱（HPLC）分析表明其为均一多糖,平均分子量为1．8万,红外光谱分析具有多糖特征吸收峰,紫外光谱未见核酸和蛋白质特征吸收峰,通过纸层析和高效液相色谱（HPLC）以及咔唑-硫酸法分析,判断其含有半乳糖、阿拉伯糖和半乳糖醛酸。总糖含量为89．3％,其中糖醛酸含量为3．36％。给小鼠腹腔注射多糖,能提高腹腔巨噬细胞的吞噬功能和吞噬指数（P〈0．01）,对移植性肿瘤S180有一定的抑制作用。     

浒苔多糖的分离,纯化和分析

浒苔经热水提取,Ｓｅｖａｇｅ法除去蛋白质,用乙醇沉淀,Ｓｅｐｈａｄｅｘ－Ｇ－１００柱层析,得浒苔多糖（简称ＥＰ）精制品。经ＳｅｐｈａｄｅｘＧ－２００柱导析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析表未见有核酸和蛋白质的特征及吸上峰,总糖含量为８８．８％,基中糖酸酸含量为３３．６％。单糖组成为Ｌ－阿拉伯糖,Ｌ－岩藻糖,Ｄ－甘露糖,Ｄ－半乳糖及Ｄ－葡萄糖。平均分子量为２５００     

桔梗多糖的提取与纯化

中药桔梗饮片经水浸提、乙醇分步沉淀,获得乙醇终浓度60%和80%的两个粗多糖组分,分别命名为PPS60和PPS80。经Molish反应、Fehling试剂反应呈阳性,碘-碘化钾反应呈阴性,说明PPS60和PPS80中均含有多糖和还原糖,但不含有淀粉。运用苯酚-硫酸法测得PPS60和PPS80的总糖含量分别为82.25%和84.17%;3,5-二硝基水杨酸（DNS）法测得还原糖含量分别为6.21%和8.05%。PPS60和PPS80经过Sevage法除蛋白、透析、浓缩和Sephrose 6B凝胶柱层析获得两个单一组分。这些结果为进一步研究桔梗多糖奠定了基础。     

硫酸酯化马尾松花粉多糖对小鼠脾脏B淋巴细胞免疫调节作用的研究

研究60％乙醇提取的马尾松花粉多糖组分D（PPM60-D）及其硫酸酯化物（SPPM60-D）对小鼠脾脏B淋巴细胞增殖、细胞内游离钙离子浓度（[Ca2＋]i）及抗体生成的影响。水煮醇沉法提取得到粗多糖,乙醇分级沉淀得到60％乙醇沉淀多糖PPM60,SephacrylS-400HR分离纯化得到多糖组分D,用氯磺酸-吡啶法对组分D进行硫酸酯化,尼龙毛法分离B淋巴细胞,MTT法测定其增殖,荧光分光光度计测定B淋巴细胞[Ca2＋]i,溶血空斑实验（PFC）和定量溶血分光光度（QHs）法测定B细胞抗体生成情况。结果显示,SPPM60-D相对于PPM60-D能更显著地提高B淋巴细胞的增殖以及[Ca2＋]i（P〈0．011：经TAK-242、LY294002、U73122、低分子肝素、维拉帕米和2-APB抑制剂作用后,均可抑制SPPM60-D和PPM60-D所致的[Ca2＋]i）升高（P〈0．05或P〈0．01）;PFC和QHS检测证实,SPPM60-D对于促进B淋巴细胞的分化及抗体的生成有显著作用,而PPM60-D的作用较弱。以上研究表明,PPM60-D经过硫酸酯化改性后,活性明显提高,推测SPPM60-D可与B淋巴细胞上TOLL样受体4（TLR4）结合,通过TLR4-P13K-PLC—IP3R信号通路使钙库释放激活的钙通道（CRAC）打开,从而使[Ca2＋]i）升高来激活B淋巴细胞,进而提高其体外增殖和抗体生成能力。     

短裙竹荪多糖Dd-S3P的分离纯化及其性质研究

短裙竹荪子实体经2％Na2CO3溶液提取,用蛋白酶法和Sevag法相结合除去蛋白,乙醇分级沉淀,级分3经DEAE－SephadexA－25柱层析纯化得到短裙竹蒸蒸荪多糖DdS3P．经测定该多糖为均一组分,分子量约为3．8×105,红外光谱呈现出典型的多糖吸收峰,含有α-型糖苷连接键,紫外扫描无核酸和蛋白质的特征吸收峰．纸层析和气相层析分析得知Dd－S3P含有D-葡萄糖（D－Glc）、D-甘露糖（D－Man）和D-木糖（D－Xyl）,其摩尔比为1．83：1．00：1．21．经动物试验表明此多糖对小鼠S-180有一定的抑制作用,抑瘤率约31．3％．     

猴头多糖HEP-2化学成分研究

猴头菌(Hericium erinaceus (Bull.) Pers．)的菌丝体培养物经水溶、乙醇沉淀、除蛋白等步骤得到猴头多糖(HEP),利用柱层析从HEP中分离得到两个组分HEP-1和HEP-2。用高效液相凝胶法(HPGPC)鉴定了HEP-2的纯度及分子量,通过有机元素分析、IR、ICP等多种分析方法确定HEP-2为一种硫酸化复合多糖,首次确定了猴头多糖的基本化学组成。     

Phylogeny of Ephemeroptera (mayflies) based on molecular evidence

This study represents the first molecular phylogeny for the Order Ephemeroptera. The analyses included 31 of the 37 families, representing approximately 24% of the genera. Fifteen families were supported as being monophyletic, five families were supported as nonmonophyletic, and 11 families were only represented by one species, and monophyly was not testable. The suborders Furcatergalia and Carapacea were supported as monophyletic while Setisura and Pisciforma were not supported as monophyletic. The superfamilies Ephemerelloidea and Caenoidea were supported as monophyletic while Baetoidea, Siphlonuroidea, Ephemeroidea, and Heptagenioidea were not. Baetidae was recovered as sister to the remaining clades. The mayfly gill to wing origin hypothesis was not supported nor refuted by these data. Mandibular tusks were supported as having at least one loss in Behningiidae and, together with the burrowing lifestyle, possibly two origins. The fishlike body form was supported as plesiomorphic for mayflies with multiple secondary losses. Topological sensitivity analysis was used as a tool to examine patterns concerning the stability of relationships across a parameter landscape, providing additional information that may not have been acquired otherwise.     

The partially degraded lipopolysaccharide of Burkholderia cepacia and ornithine-containing lipids derived from some Gram-negative bacteria are useful complex lipid adjuvants

The partially degraded lipopolysaccharide of Burkholderia cepacia (LPSdegr) and the ornithine-containing lipids were purified from some bacteria. The substances were developed as complex lipid adjuvants, because they have weak toxicity and are able to activate the immune systems of the living body. After various toxoid antigens such as pertussis toxoid, diphtheria toxoid and tetanus toxoid were mixed with the complex lipid adjuvants, the mixtures were administered to mice subcutaneously. Antitoxoid IgG antibody titers in the serum were measured several times over 3 months. The efficacy of the LPSdegr as adjuvant was almost as high as that of the ornithine-containing lipids, and it was almost equal to that of the aluminum hydroxide adjuvant (Alum), which is generally used as a vaccine adjuvant.     

新疆天山雪莲体胚诱导与分化研究

以新疆天山雪莲的叶片为外植体,分别用不同配方培养基诱导愈伤组织,后进行体胚诱导和分化培养形成再生雪莲植株.结果表明,诱导愈伤组织的最适培养基为MS 2,4-D 0.5 mg/L BA 1.5 mg/L,诱导率可达到100%;愈伤组织转移至MS 2,4-D 0.5 mg/L BA 1.5 mg/L培养基进行继代培养,增殖后的愈伤组织转移到MS 2,4-D 0.2 mg/L的液体培养基后成功诱导出雪莲体胚,出胚率达40%;将体胚接至MS ABA 0.5 mg/L培养基后,结果分化生长出大量的再生雪莲幼苗.     

The action of beta-galactosidase (Escherichia coli) on allolactose.

The parameters involved in the action of beta-galactosidase (EC 3.2.1.23) (Escherichia coli) on allolactose, the natural inducer of lac operon in E. coli, were studied. At low allolactose concentrations only galactose and glucose were formed, while at high allolactose concentrations transgalactolytic oligosaccharides were also produced. Detectable amounts of lactose were not formed. The V and Km values (49.6 U/mg and 0.00120 M, respectively) indicated that allolactose is as good if not a better substrate of beta-galactosidase as lactose. The pH optimum with allolactose (7.8-7.9) as well as its activation by K+ (as compared to activation by Na+) were similar to the case with lactose as substrate. The alpha-anomer of allolactose was hydrolyzed about two times as rapidly as was the beta-anomer.     

Computational simulation of the statistical properties of unfolded proteins

A simple Monte Carlo method was used to generate ensembles of simulated polypeptide conformations that are restricted only by steric repulsion. The models used for these simulations were based on the sequences of four real proteins, ranging in size from 26 to 268 amino acid residues, and included all non-hydrogen atoms. Two sets of calculations were performed, one that included only intra-residue steric repulsion terms and those between adjacent residues, and one that included repulsion terms between all possible atom pairs, so as to explicitly account for the excluded volume effect. Excluded volume was found to increase the average radius of gyration of the chains by 20-40%, with the expansion factor increasing with chain length. Contrary to recent suggestions, however, the excluded volume effect did not greatly restrict the distribution of dihedral angles or favor native-like topologies. The average dimensions of the ensembles calculated with excluded volume were consistent with those measured experimentally for unfolded proteins of similar sizes under denaturing conditions, without introducing any adjustable scaling factor. The simulations also reproduced experimentally determined effective concentrations for the formation of disulfide bonds in reduced and unfolded proteins. The statistically generated ensembles included significant numbers of conformations that were nearly as compact as the corresponding native proteins, as well as many that were as accessible to solvent as a fully extended chain. On the other hand, conformations with as much buried surface area as the native proteins were very rare, as were highly extended conformations. These results suggest that the overall properties of unfolded proteins can be usefully described by a random coil model and that an unfolded polypeptide can undergo significant collapse while losing only a relatively small fraction of its conformational entropy.     

化橘红黄酮类化合物抑制亚硝化反应活性研究

根据超声波最佳提取工艺条件提取得到化橘红总黄酮,经乙酸乙酯萃取分为酯溶性和水溶性两部分,比较它们对亚硝化反应的抑制作用,并分别采用硅胶柱层析对酯溶性成分分离纯化,采用大孔吸附树脂法对水溶性成分进行分离纯化,得到4个主要的黄酮类化合物,研究它们对亚硝化反应的抑制作用,以期得到抑制亚硝化活性较强的化合物。结果表明化橘红总黄酮提取率可达26.42%,酯溶性和水溶性部分均能阻断亚硝胺的合成及清除亚硝酸盐,其中水溶性黄酮提取物作用较强,分离得到的4个黄酮类化合物中柚皮苷的抑制作用较强,对亚硝胺的合成的最大阻断率可达94.7%,对亚硝酸盐的最大清除率可达92.3%。     

Growth and enzyme synthesis during continuous culture of Trichosporon cutaneum on phenol

The soil yeast Trichosporon cutaneum was grown in continuous culture with phenol as the sole carbon source. The cultures were operated as carbon-limited chemostats or as steady-state continuous cultures without carbon limitation. Selected comparative runs were also conducted on glucose or acetate as carbon source. In addition to growth parameters, the activities of several intracellular enzymes were determined, comprising those directly involved in the degradation of phenol as well as auxiliary enzymes required for the generation of reducing power. All enzymes were assayed in detergent-permea-bilized cells. Phenol was found to serve as an excellent carbon source, comparable to glucose or acetate. The utilization of phenol in T. cutaneum is very efficient as indicated by a low maintenance requirement (0.01 g phenol/g cells.h). The cell yields obtained were on the order of 0.8 g cells/g phenol. Although the phenol-limited chemostats were run with fully phenol-induced cells, a further increase in the activities of isocitrate DH(NADP(+)), maleate DH and the phenol-degrading enzymes occurred after transition to nonlimiting condition. Enzyme activities increased in parallel with increasing phenol levels in the effluent, as well as with increasing toxicity. The significance of this phenomenon is discussed. The significance of this phenomenon is discussed. This elevation in enzyme activities in not related to an increase in specific growth rate.     

Evaluation of 100% rapid rescreening of negative cervical smears as a quality assurance measure

OBJECTIVE: The objective of this study was to compare the performance of 100% rapid rescreening, 10% random rescreening and the review of smears selected on the basis of clinical criteria, as a method of internal quality control of cervical smears classified as negative during routine screening. METHODS: A total of 3149 smears were analysed, 173 of which were classified as positive and 2887 as negative, while 89 smears were considered unsatisfactory. The smears classified as negative were submitted to 100% rapid rescreening, 10% random rescreening, and rescreening based on clinical criteria. The rescreening stages were blinded and results were classified according to the Bethesda 2001 terminology. Six cytologists participated in this study, two of whom were responsible for routine screening while the other four alternated in carrying out rescreening so that no individual reviewed the same slide more than once. RESULTS: The 100% rapid rescreening method identified 92 suspect smears, of which 42 were considered positive at final diagnosis. Of the 289 smears submitted to the 10% rescreening method, four were considered abnormal but only one was confirmed positive in the final diagnosis. Of the 690 smears rescreened on the basis of clinical criteria, 10 were considered abnormal and eight received a positive final diagnosis. CONCLUSIONS: The 100% rapid rescreening method is more efficient at detecting false-negative results than 10% random rescreening or rescreening on the basis of clinical criteria, and is recommended as an internal quality control method.     

Crocin prevents metabolic syndrome in rats via enhancing PPAR-gamma and AMPK

Metabolic syndrome (Mets) is a major health hazard. The syndrome is strongly linked to cardiovascular disease. The objective of the current study was to address whether or not crocin could protect against experimentally-induced MetS in rats as well as the possible underlying mechanisms. Animals were allocated into 4 groups. The first one served as control and was kept on regular food pellets and drinking water. The other three groups were subjected to experimental MetS. Induction of MetS was achieved by keeping rats on food pellets containing 3% NaCl; and drinking water enriched with 10% fructose. The first and second groups were daily gavaged with saline. The third and fourth groups were daily administered crocin 5 and 10 mg/kg, respectively. The treatment continued for 12 consecutive weeks. Crocin significantly reduced body weight gain and adiposity index as compared to MetS group. Also, crocin protected against the occurrence of hyperglycemia and insulin resistance as indicated by controlled values of HOMA-IR. Crocin protected against MetS-induced dyslipidemia as well as the rise blood pressure. These beneficial effects were accompanied by enhanced serum levels of PPARγ & AMPK and inhibited serum levels of IL-6 and TNF-α. In conclusion, crocin protects against experimentally-induced MetS. This can be attributed, at least partly, to activation of PPARγ and AMPK as well as inhibiting inflammation.     

A cytogenetic study of men environmentally and occupationally exposed to airborne pollutants.

The level of sister-chromatid exchanges (SCE), high-frequency cells (HFC), chromosomal aberrations (CA) as well as the proliferation rate index (PRI) were measured in peripheral blood lymphocytes from three groups of volunteers. The environmentally exposed donors were residents from the vicinity of a coke factory; the occupationally exposed persons were cokery workers, while rural region inhabitants served as a control group. Compared with the control group, statistically significant increases of SCE and HFC, as well as decreased cell kinetics (PRI) were observed for both occupationally and environmentally exposed groups. The effect was especially pronounced when only smokers were taken into account. A statistically significant increase of CA was observed in the environmentally exposed group when CA including gaps (CA + G) were evaluated. The proportion of HFC was found to be the most sensitive method to detect genetic effects on the tested human population. This study demonstrates the usefulness of all 4 biomarkers (SCE, HFC, CA and PRI) in monitoring populations exposed to ambient pollution and clearly indicates effects from residential as well as occupational exposure to industrial air pollutants.