毛尖蘑子实体粗多糖对肿瘤生长及免疫器官的影响

目的 探讨毛尖蘑子实体粗多糖(MJMP)对H22荷瘤小鼠的抗肿瘤作用以及对免疫器官的影响.方法 将不同浓度的MJMP作用于H22荷瘤小鼠,计算胸腺指数、脾指数、抑瘤率,观察H22荷瘤小鼠免疫器官的变化及肿瘤生长情况.结果 多糖作用组与空白对照组及阴性对照组脾指数比较无统计学意义(P＞0.05),胸腺指数有差异(P＜0.05).小鼠体内实验显示MJMP可明显抑制肿瘤生长.结论 MJMP对小鼠免疫功能有增强的作用,对肿瘤的生长具有抑制作用.     

松杉灵芝子实体抗肿瘤活性多糖的提取、分离与鉴定

目的 研究松杉灵芝子实体中抗肿瘤活性多糖的提取、分离与鉴定的方法.方法 把依次用热水、1%草酸铵、5%氢氧化钠提取得到的粗多糖经过离子交换层析、凝胶过滤层析及亲和层析分离精制,将得到的多种活性多糖进行小白鼠S180抗肿瘤实验,并分别测定多种活性多糖的分子量、构成糖组成、蛋白含量、氨基酸组成等.结果 分离精制得到的多种活性多糖中,有6种多糖具有比较明显的抗肿瘤药理活性.结论 松杉灵芝子实体中含有的多种活性多糖具有明显的抗肿瘤药理活性,作为东北长白山地区名贵的民间药物可广泛开发利用.     

鳖甲多糖对小鼠抗肿瘤作用及其机理的研究

目的研究鳖甲多糖对S180荷瘤小鼠抗肿瘤的免疫调节作用。方法制备S180荷瘤小鼠动物模型，随机分为荷瘤对照组（Ⅰ）、低剂量组（Ⅱ）、中剂量组（Ⅲ）、高剂量组（Ⅳ）；通过测定荷瘤小鼠的瘤重、抑瘤率及瘤体比观察鳖甲多糖对肿瘤生长的影响；通过检测脾细胞的增殖分化、NK细胞的活性以及腹腔巨噬细胞的吞噬功能，研究鳖甲多糖对荷瘤小鼠非特异性免疫和细胞免疫功能的影响。结果鳖甲多糖能明显减小S180荷瘤小鼠的瘤重和瘤体比（P〈0．05），能明显抑制肿瘤的生长（P〈0．05），Ⅱ、Ⅲ、Ⅳ组的平均抑瘤率分别为30％、37％、45％；鳖甲多糖能明显增强S180荷瘤小鼠腹腔巨噬细胞的吞噬功能（P〈0．05）；鳖甲多糖能明显提高S180荷瘤小鼠脾细胞的转化功能和NK细胞的活性（P〈0．05）。结论鳖甲多糖能明显抑制S180荷瘤小鼠肿瘤的生长，其作用机制可能是通过增强荷瘤小鼠的非特异性免疫功能和细胞免疫功能。     

胡颓子多糖对小肠辐射损伤的保护作用

目的:观察长叶胡颓子多糖对小肠辐射损伤的的保护作用及其机制.方法:昆明小鼠60只随机分为:正常组、辐射组、不同剂量(100,300,500 mg/kg)胡颓子多糖给药组.采用~(60)Coγ射线一次性全身均匀照射建立小肠辐射损伤模型,相应灌胃用药2 wk后,测定小鼠肠段的肠腺存活率百分比和二胺氧化酶(DAO)含量,检测肠黏膜蛋白质和DNA含量(mg/cm),肠组织中SOD活性和MDA含量,血清中TNF、IL-4、IL-6水平.结果:不同剂量长叶胡颓子多糖灌胃均不同程度提高十二指肠、空肠和回肠段肠腺存活率(P<0.05),促进肠黏膜蛋白质、DNA合成(P<0.05或P<0.01),降低血液中TNF水平(6.89±1.33,6.74±1.24,5.87±1.13 vs 8.53±1.08;P<0.05或P<0.01)和MDA含量,提高IL-4(9.79±2.23,10.83±2.34,11.36±2.51 vs 7.40±2.08;P<0.05或P<0.01)、IL-6水平(49.36±9.65,55.26±11.32,59.68±13.65 vs 43.74±12.91;P<0.05或P<0.01)和SOD活性,且与用药剂量呈量效关系.结论:长叶胡颓子多糖通过促进肠黏膜蛋白质和DNA合成、增强抗氧化能力、调节细胞因子异常和损伤修复作用维护肠黏膜的完整性、增强屏障作用,有效减轻小肠辐射损伤.     

微生物多糖提取、分离和纯化方法的技术策略

<正>多糖广泛分布于高等植物、地衣、海藻、动物和微生物中,具有提高免疫力、降血糖、抗肿瘤、抗病毒等多种生物活性,针对一些常见的老年性急慢性疾病具有预防和治疗作用。由于微生物具有繁殖能力强、周期短、成本低、易控制的特点,所以普遍采用微生物发酵法生产多糖,其产量及质量稳步提高,性价比也优于动植物来源的多糖。目前,在食品工业中,微生物多糖已作为胶凝剂、成膜剂、保鲜剂、乳化剂等添加剂,广泛应用于老年保健食品,还有一些微生物胞外多糖及其衍生物具有     

毛尖蘑子实体粗多糖对H22荷瘤小鼠的抑瘤作用

目的探讨毛尖蘑子实体粗多糖(MJMP)对H22荷瘤小鼠的抑瘤作用和可能机制。方法建立小鼠H22实体瘤模型,并对小鼠H22实体瘤模型进行MJMP处理,HE染色观察凋亡细胞形态,RT-PCR法分析Caspase-3 mRNA表达水平。结果小鼠体内实验显示MJMP可明显抑制肿瘤生长。HE染色观察到凋亡细胞的形态学改变,RT-PCR表明Caspase-3 mRNA基因表达均增强。结论 MJMP对H22肿瘤具有明显抑制作用,其分子机制可能与Caspase-3基因表达上调有关。     

树舌多糖GF注射液对化疗药物的增效减毒作用及其机制研究

目的探讨树舌多糖GF注射液对化疗药物(CTX)的增效减毒作用及其作用机制。方法采用移植性H22肝癌小鼠，随机分为阴性对照组、CTX组和联合用药组。连续给药10d后，测定其抑瘤率、WBC数、吞噬指数和IL-2活性。结果树舌多糖GF注射液能提高CTX的抑瘤率，能减轻CTX的毒副作用，并能提高单核吞噬细胞功能和IL-2活性。结论树舌多糖GF注射液对化疗药物CTX具有明显的增效减毒作用，可能是通过增强机体的免疫功能实现的。     

十全大补冲剂对喃呋啶抑瘤活性的增强作用及其毒副反应的药效学研究

本文研究了十全补冲剂对喃呋啶抑瘤活性的增强作用及其毒副反应的影响。结果表明，不同剂量的十全大补冲剂与喃呋啶伍用均能明显提高喃呋啶对小鼠移植肿瘤Ｓ１８０、Ｌｅｗｉｓ及Ｈ２２三种瘤株的抑瘤率；与单用喃呋啶相比Ｐ值＜０．０５～０．０１；对喃呋啶所致小鼠白细胞下降、免疫器官萎缩、巨噬细胞吞噬功能降低、血清溶血素形成减少等均有显著保护作用，同时还能提高ＮＫ细胞活性     

板党多糖的抗衰老作用及机制

目的探讨板党多糖抗衰老作用及其机制。方法复制臭氧(O3)损伤小鼠衰老模型,将60只昆明种小鼠随机分6组(n=10):空白组,模型组,香菇菌多糖组和板党多糖低、中、高剂量组,行板党多糖灌胃20 d。测定胸腺和脾脏指数,取血、肝、脑测定丙二醛(MDA)、氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)。结果板党多糖能显著提高衰老小鼠胸腺指数和脾脏指数;显著降低MAD含量并提高SOD、GSH-Px和CAT活力。结论板党多糖可通过增强机体免疫功能、提高机体抗氧化能力和直接清除自由基发挥其延缓衰老的作用。     

黄芪多糖抗动脉粥样硬化作用及其机制的研究

目的利用高脂饲料诱发动脉粥样硬化(As)模型,观察黄芪多糖(APS)对As模型的影响,探讨该药的治疗作用及可能机制。方法将30只健康国产雄性家兔,随机分为3组:正常对照组、模型组、黄芪多糖治疗组,每组10只。对照组给予正常颗料饲料,其余2组从实验第1天起给予高脂饲料(80%基础饲料中加入15%蛋黄粉、0.5%胆固醇和5%猪油),用高脂饲料建立家兔动脉粥样硬化模型。黄芪多糖组每天腹腔注射黄芪多糖500 mg/kg;对照组、模型组给予等体积的生理盐水,体积为4 mL/kg,共给药50天。末次给药后24 h,从上腔静脉取血后处死动物。腹主动脉形态学变化在光镜下观察,并测定家兔血清中总胆固醇(TC)、甘油三酯(TG)、一氧化氮(NO)、内皮素1(ET-1)、超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化活力(T-AOC)的变化。结果模型组与对照组比较,总胆固醇(TC)、甘油三酯(TG)、丙二醛(MDA)、内皮素1(ET-1)明显升高(P<0、O1),一氧化氮(NO)、SOD及总抗氧化活力(T-AOC)明显下降(P<0、O1),主动脉内膜粥样斑块面积较大;而APS组与模型组比较,总胆固醇(TC)、甘油三酯(TG)...     

Influence of cyclic nucleotides on receptor binding, immunological activity, and microheterogeneity of human choriogonadotropin synthesized in placental tissue culture

The regulation of the biosynthesis of choriogonadotropin (hCG) in tissue culture by human first trimester placenta in the presence of the following cyclic nucleotides and 3-isobutyl-1-methylxanthine (IBMX) was studied (concentrations in parentheses): IBMX (0.1 mM), cAMP (1 mM) + IBMX (0.1 mM), cGMP (0.1 mM) + IBMX (0.1 mM), 8-bromo-cAMP (0.5 mM) and 8-bromo-cGMP (0.5 mM). The medium concentration of hCG follows an optimum curve at all conditions, showing highest values at day 3 of the culture. The efficacy of the substances to cause an increase in the hCG medium concentration was in the following order: IBMX less than control less than 8-bromo-cGMP = 8-bromo-cAMP less than cGMP + IBMX less than cAMP + IBMX. The synthesized hCG was examined with respect to its receptor binding activity (LH/hCG receptor of rat testes), the activities to stimulate adenylate cyclase as well as testosterone biosynthesis in purified mouse Leydig cells, the immunological activity, and the microheterogeneity in isoelectric focusing. Only in the presence of 8-bromo-cAMP, 8-bromo-cGMP, and cGMP + IBMX was hCG synthesized, which differs significantly in the investigated properties from hCG of the control cultures. Only in the presence of 8-bromo-cGMP is the ratio of receptor binding activity/immunological activity optimal (near 1). In the presence of both 8-bromo-cAMP and 8-bromo-cGMP, microheterogeneity of hCG in isoelectric focusing was diminished and the synthesis of more acidic hCG subpopulations was favoured.     

Biological and immunological characterization of human luteinizing hormone: II. A comparison of the immunological and biological activities of pituitary extracts after electrofocusing using different standard preparations

The profile of immunologically active human luteinizing hormone (hLH) was determined in aqueous pituitary extracts after electrofocusing using two radioimmunoassay systems and the estimates for each fraction were compared to those obtained by an in vitro bioassay method. Similar biological and immunological profiles were obtained in the pH 7.0-9.0 region, where most of the biological activity was present. Biological to immunological (B/I) ratios ranging from 0.6 to 1.7 (mean ratio 1.01; n = 21) were found in the major biologically active fractions of this pH region when a highly purified human pituitary LH preparation (68/40) was used as standard in both types of assays. The close proximity of these ratios to unity indicates a similar composition of biological and immunological activities in all these fractions in relation to that of the highly purified standard. However, marked discrepancies were observed in the pH region 3.0-7.0 where the B/I ratios ranged from 0.1 to 0.9, indicating the presence of immunological activity associated with relatively little biological activity. When impure human LH preparations of pituitary (69/104) and urinary (hMG 2nd IRP) origin were used as standards for the bioassay and radioimmunoassay of the hLH present in the major fractions of the pH region 7.0-9.0, significantly higher B/I ratios were obtained than with the use of the highly purified standard (68/40). These elevated B/I ratios are attributed to the presence in the impure standard preparations of immunological activity, which is associated with little or no biological activity. These observations may provide an explanation for the differences in B/I ratios which were reported for hLH in plasma, using different standard preparations.     

当归多糖对免疫性肝损伤小鼠肝中一氧化氮合酶及凋亡相关基因的影响

目的研究免疫性肝损伤中结构型（ｃＮＯＳ）、诱导型（ｉＮＯＳ）一氧化氮合酶及凋亡相关基因Ｂａｘ、Ｂｃｌ－２表达的变化,在进一步探讨免疫性肝损伤机制的同时观察当归多糖的干预调控作用。方法建立卡介苗和脂多糖诱导的小鼠免疫性肝损伤模型,给予当归多糖 ３０ ｍｇ／ｋｇ、 ６０ ｍｇ／ｋｇ,测定血中 ＡＬＴ、 ＧＳＴ活性及肝中 ＮＯ含量;用免疫组织化学方法观察ｃＮＯＳ、ｉＮＯＳ、Ｂａｘ、Ｂｃｌ－２的表达。结果免疫性肝损伤小鼠ｓＡＬＴ、ｓＧＳＴ及ＮＯ含量明显升高,ｉＮＯＳ含量为正常鼠的１７．８倍,ｃＮＯＳ无明显变化,抗调亡基因Ｂｃｌ－２呈阴性表达,而具有启动凋亡信号、抑制Ｂｃｌ－２表达的 Ｂａｘ增加３１．１％。小剂量当归多糖可使 ｓＡＬＴ、 ｓＧＳＴ及 ＮＯ含量分别降低 ２４．６％、 ４０．８％、 １８．４％, ｉＮＯＳ、Ｂａｘ表达下降８４．２％、 ３７．１％,并使ｃＮＯＳ表达升高６６．８％, Ｂｃｌ－２表达增加 ３．３８倍;大剂量当归多糖可使ｓＡＬＴ、 ｓＧＳＴ及ＮＯ含量分别降低３６．６％、３４．５％、１６、９％,对Ｂａｘ表达的降低作用及对ｃＮＯＳ、Ｂｃｌ－２表达的增加作用不及小剂量明显,但能明显地降低ｉＮＯＳ的表达。结...     

重组日本血吸虫特异性IgE相关蛋白的纯化及免疫学活性鉴定

目的　纯化重组日本血吸虫特异性IgE抗体相关蛋白和鉴定其免疫原性。　方法　大量表达Sj43B/pGEX 6p 1 /BL2 1重组克隆菌 ,超声粉碎后离心获得融合表达的重组蛋白包涵体。经TNMFX缓冲液分步洗涤后 ,将包涵体溶解液经FPLC分离 ,获得重组融合蛋白组分 ,再经巯基二硫键转换复性后 ,用于免疫小鼠获得抗血清 ,分别用dot ELISA法和Westernblotting法对融合蛋白引起的特异性血清抗体同型反应进行鉴定。　 结果　分步洗涤可有效去除重组蛋白包涵体沉淀中混杂的多数杂蛋白成分 ,FPLC分离可获得高纯度重组蛋白。用复性后的重组融合蛋白免疫小鼠 ,其中目的蛋白可引起特异性IgE抗体反应 ,而担体蛋白 2 6kDaGST不引起特异性IgE应答 ,可引起特异性IgG抗体反应 ,目的蛋白则否。　结论　重组质粒Sj43B/pGEX 6p 1表达的融合蛋白 ,其目的蛋白部分免疫小鼠可产生特异性IgE抗体。     

屋尘螨8类变应原(Der p 8)克隆表达、纯化及免疫原性鉴定

目的 对屋尘螨(Dermatophagoides pteronyssinus) 8类变应原Der p 8进行克隆表达、纯化及免疫原性分析,并用生物信息学方法分析其同源性。方法 根据已知Der p 8基因序列,设计PCR引物;提取屋尘螨总RNA,逆转录聚合酶链式反应(RT-PCR)扩增得到Der p 8的cDNA片段,以cDNA和设计的引物进行PCR扩增;扩增产物连接至T载体并转化入大肠埃希菌(E.coli Top10)中,培养后挑选阳性单克隆菌落测序。测序正确后,转入表达载体pET-32a,经酶切鉴定、测序后转入表达菌BL21,大量表达重组蛋白Der p 8。用亲和层析法纯化后进行免疫印迹分析(Western blotting)。利用生物信息学方法分析Der p 8基因的同源性。 结-果 酶切鉴定显示Der p 8表达载体构建成功,目的基因分子量约为700 bp;SDS-PAGE电泳显示Der p8重组蛋白成功表达,分子量约为 38 kD,且主要以包涵体形式存在。Western blotting结果显示Der p 8重组蛋白具有免疫原性。同源性分析结果显示本实验克隆Der p 8基因与NCBI基因库的Der p 8基因同源性为76.97%。结论 本实验成功克隆表达并纯化出具有免疫原性的Der p 8重组蛋白,为临床上应用重组蛋白于诊断和治疗提供理论基础。     

Larvicidal activity of Wrightia tinctoria R. BR. (Apocynaceae) fruit and leaf extracts against the filarial vector Culex quinquefasciatus Say (Diptera: Culicidae)

ObjectiveTo determine the larvicidal activity of crude aqueous and petroleum ether extracts of Wrightia tinctoria fruits and leaves against the filarial vector, Culex quinquefasciatus.MethodsThe larvicidal activity was evaluated at concentrations of 0.06%, 0.12%, 0.25%, 0.50% and 1.00%. Larval mortality was observed for 24 and 48 h.ResultsAmong the plant parts tested, aqueous fruit extract exhibited highest larvicidal activity followed by aqueous leaf extract with LC₅₀ values of 0.17% and 0.09%; 0.21% and 0.11% after 24 and 48 h respectively.ConclusionsFurther investigations are needed to elucidate this activity against a wide range of all stages of mosquito species and also the active ingredient(s) of the extract responsible for larvicidal activity should be identified.     

Establishment and application of an experimental model of human fetal hepatocytes for investigation of the protective effects of silybin and polyporus umbellalus polysaccharides

AIM: To establish a new experimental model system of human fetal hepatocytes to study the mechanisms underlying the protective effect of silybin and polyporus umbellalus polysaccharides (PSP) on the cellular ultrastructure.METHODS: Human fetal hepatocytes were obtained from the liver of a human fetus that resulted from a medically necessary induced labor; the mother provided informed consent for sampling, experimental use and publication of findings. The hepatocytes were cultured and then pretreated with silybin or PSP or without either (control), after which the treated cells were exposed to CCl₄ for 4 h. Changes in cellular ultrastructure were observed by scanning electron microscopy and transmission electron microscopy, and changes in alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD) were assayed.RESULTS: Levels of ALT and AST were significantly decreased, and level of SOD was elevated in the two pretreatment groups following CCl₄ exposure, as compared to the control group. The cellular integrity and ultrastructure were well preserved in the two pretreatment groups but were seriously damaged in the control group.CONCLUSION: The CCl₄-induced hepatotoxic cell model system of human fetal hepatocytes is an effective tool for studying the hepatoprotective effect of drugs and may be applicable for studies to screen medicines for treatment of hepatitis.     

第二届全国人工肝及血液净化学术年会会议纪要

由中华医学会肝病学分会重肝与人工肝学组、中国肝炎防治基金会人工肝及血液净化科技攻关基金委员会、全国人工肝及血液净化攻关协作组联合主办、大连市第六人民医院协办的第二届全国人工肝及血液净化学术年会于2005年9月1日至4日在辽宁大连举行。会议共收录论文228篇，来自中国、美国、德国、日本的700余名代表参加本次会议。卫生部、科技部、中华医学会、中华医院管理学会、大连市的有关领导出席了会议。国内外肝病内外科、传染病、消化、血液净化、肾病、生物医学工程等领域的多位知名专家主持会议并做了专题发言。大会开设的自由论坛还使每位代表都有机会发言、交流，会场气氛热烈。会议就重型肝炎肝衰竭的机制、诊断、内科药物治疗、人工肝、血液净化、肝移植、细胞移植等方面进行了广泛、深入的探讨与交流。会议期间还举行了《实用人工肝及血液净化操作手册》发行仪式，中国肝炎防治基金会人工肝及血液净化科技攻关基金共资助7项2006-2007年度人工肝及血液净化科研课题。与会专家介绍了有关重型肝炎、人工肝及血液净化技术的多项最新进展。     

Total body and serum electrolyte composition in heart failure: the effects of captopril

We compared the long-term effects of captopril and placebo onpatients with heart failure in a double blind crossover fashion.Serum and total body elecrolytes were measured and the responseto 6 week periods of treatment with captopril determined. Duringthe placebo phase of the study, total body potassium was lowat 92±14% of predicted normal (P<0.05) and total bodysodium was high at 104+7% of predicted normal (P<0.05). Totalbody chlorine did not differ from predicted normal (99+12%).In those patients with active plasma renin concentrations abovethe normal range (greater than 50 µU ml^–1) totalbody potassium was even more markedly deplete (85+13% of predictednormal). This group was also characterized by lower serum potassiumand sodium concentrations and lower blood pressure. Total body potassium increased significantly on captopril, andthe rise was greatest in those with the highest plasma reninconcentrations during the placebo phase of the study. However,captopril had no significant effect on total body sodium andchlorine or weight indicating that no long-term natriuresishad occurred.     

重组人成骨蛋白-1的纯化及其生物学活性研究

目的 获得高纯度的重组人成骨蛋白-1（rhOP-1）,探讨大肠杆菌表达rhOP-1的生物学活性。方法 表达rhOP-1工程菌经发酵后,裂菌、提取包涵体,层析法纯化蛋白并逐步降低尿素浓度透析复性。采用改良MTT法检测rhOP-1对NIH3T3细胞增殖和对硝基苯酚法（PNPP）检测rhOP-1对细胞碱性磷酸酶（ALP）活性的影响,同时在体内通过小鼠肌袋埋植实验测定rhOP-1活性。结果 经纯化的rhOP-1纯度可达98％。体外rhOP-1浓度在0．06～6μg／ml可明显促进细胞生长和ALP活性;体内rhOP-1埋植剂植入小鼠肌间隙2周后,可明显提高体内钙含量,且成骨趋势明显。结论 原核表达的rhOP-1具有良好的生物学活性。